Systems, methods, and compositions for rna-guided rna-targeting crispr effectors

ABSTRACT

This disclosure provides systems, methods, and compositions for RNA-guided RNA-targeting CRISPR effectors for the treatment of diseases, and for use as diagnostics. In addition, nucleotide deaminase functionalized CRISPR systems for RNA editing RNA knockdown, viral resistance, splicing modulation, RNA tracking, translation modulation, and epi-transcriptomic modifications are disclosed.

CROSS REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional PatentApplication Ser. No. 63/365,281, filed May 25, 2022. The entire contentsof the above-referenced patent application is incorporated by referencein its entirety herein.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under AI149694 andHG011857 awarded by National Institutes of Health. The government hascertain rights in the invention.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in .xml format and is hereby incorporated byreference in its entirety. The .xml copy, created on May 18, 2023, isnamed 739749083474-034 and is 341,432 bytes in size.

TECHNOLOGY FIELD

The subject matter disclosed herein is generally related to systems,methods, and compositions for RNA-guided RNA-targeting CRISPR effectorsfor the treatment of diseases and diagnostics.

BACKGROUND

RNA-targeting tools are important for studying RNA biology, forengineering genes, and for developing RNA therapeutics, among others.These tools can regulate intracellular and intercellular target-genefunction and expression as well as manipulate specific target-genomicinformation. Few RNA-targeting tools have been developed, and those thathave can present challenges. For instance, some have weak activity inmammalian cells, and some have collateral effects, which can be toxic incertain cell types. In some circumstances, the size of the RNA-targetingtools can be a barrier to their use. Delivery of the RNA-targeting toolscan also be difficult. Thus, there remains a need for effective toolsfor RNA-targeting tools and their delivery.

SUMMARY

The present disclosure provides compositions and systems for RNA-guidedRNA-targeting CRISPR effectors for the treatment of diseases anddiagnostics.

In one aspect, provided herein is a polypeptide comprising an amino acidsequence at least 85% identical to the amino acid sequence of any one ofSEQ ID NOs: 1-4. The amino acid sequence of the polypeptide comprises atleast one amino acid modification or mutation relative to the amino acidsequence of SEQ ID NO: 1-4.

In some embodiments, the amino acid sequence of the polypeptide is atleast 85%, at least 90%, at least 95%, or at least 99% identical to theamino acid sequence of SEQ ID NO: 1-4.

In some embodiments, the amino acid sequence of the polypeptidecomprises the amino acid sequence of SEQ ID NO: 1-4.

In some embodiments, the at least one amino acid modification ormutation comprises: removing an amino acid; adding an amino acid;replacing an amino acid with no charge with an amino acid with apositive charge; or replacing an amino acid with a negative charge withan amino acid with a positive charge.

In some embodiments, the amino acid without charge is selected from thegroup consisting of serine, threonine, asparagine, glutamine, cysteine,glycine, proline, alanine, valine, isoleucine, leucine, methionine,phenylalanine, tyrosine, and tryptophan.

In some embodiments, the amino acid with a negative charge is selectedfrom the group consisting of aspartic acid and glutamic acid.

In some embodiments, the amino acid with a positive charge is selectedfrom the group consisting of arginine, histidine, and lysine.

In some embodiments, the amino acid sequence of the polypeptidecomprises 1, 2, 3, or 4 amino acid modifications or mutations.

In some embodiments, the amino acid sequence of the polypeptidecomprises an alanine at a position corresponding to position 43 of SEQID NO: 1; an alanine at a position corresponding to position 55 of SEQID NO: 55; and/or an alanine at a position corresponding to position 152of SEQ ID NO: 1.

In some embodiments, the polypeptide comprises a deletion of one or moreamino acid residues at positions 979 through 1293 of SEQ ID NO: 1; atpositions 1007 through 1220 of SEQ ID NO: 1; and/or at positions 1146through 1211 of SEQ ID NO: 1.

In another aspect, provided herein is a composition that cleaves an RNAtarget comprising a guide RNA that specifically hybridizes to the RNAtarget and a polypeptide.

In some embodiments, the guide RNA comprises a mismatch distance that isabout 20-65% of the length of the guide.

In some embodiments, the guide RNA has a sequence with a length of fromabout 20 to about 53 nucleotides (nt), optionally from about 25 to about53 nt, more optionally from about 29 to about 53 nt, or optionally fromabout 40 to about 50 nt.

In some embodiments, the guide RNA is a pre-crRNA.

In some embodiments, the guide RNA is a mature crRNA.

In some embodiments, the RNA target is a single-strand RNA (ssRNA).

In some embodiments, the RNA target is in a cell.

In some embodiments, the cell is a prokaryotic cell.

In some embodiments, the cell is a eukaryotic cell.

In some embodiments, the eukaryotic cell is a mammalian cell.

In some embodiments, the mammalian cell is a human cell.

In some embodiments, the guide RNA comprises a mismatch that is about 20to about 30 nucleotides from a non-pairing C of the guide RNA.

In another aspect, provided herein is a nucleic acid molecule encoding apolypeptide.

In some embodiments, the nucleic acid molecule encodes the guide RNA.

In some embodiments, the nucleic acid molecule further comprises anucleic acid molecule that encodes the guide RNA.

In another aspect, provided herein is a vector comprising the nucleicacid molecule.

In some embodiments, the vector is a viral vector.

In some embodiments, the viral vector is a lenti-associated viralvector, baculo-associated viral vector, or adeno-associated viralvector.

In some embodiments, the viral vector is derived from a virus selectedfrom the group consisting of Myoviridae, Siphoviridae, Podoviridae,Corticoviridae, Lipothrixviridae, Poxviridae, Iridoviridae,Adenoviridae, Polyomaviridae, Papillomaviridae, Mimiviridae,Pandoravirusa, Salterprovirusa, Inoviridae, Microviridae, Parvoviridae,Circoviridae, Hepadnaviridae, Caulimoviridae, Retroviridae,Cystoviridae, Reoviridae, Bimaviridae, Totiviridae, Partitiviridae,Filoviridae, Orthomyxoviridae, Deltavirusa, Leviviridae, Picomaviridae,Mamaviridae, Secoviridae, Potyviridae, Caliciviridae, Hepeviridae,Astroviridae, Nodaviridae, Tetraviridae, Luteoviridae, Tombusviridae,Coronaviridae, Arteriviridae, Flaviviridae, Togaviridae, Virgaviridae,Bromoviridae, Tymoviridae, Alphaflexiviridae, Sobemovirusa, Idaeovirusa,and Herpesviridae.

In another aspect, provided herein is a cell comprising a polypeptide, acomposition, a nucleic acid molecule, and/or a vector.

In some embodiments, the cell is a prokaryotic cell.

In some embodiments, the cell is a eukaryotic cell.

In some embodiments, the eukaryotic cell is a mammalian cell.

In some embodiments, the mammalian cell is a human cell.

In another aspect, provided herein is a method of cleaving an RNA targetin a cell comprising providing to the cell a polypeptide, a composition,a nucleic acid molecule, and/or a vector.

In another aspect, provided herein is a method of stabilizing an RNAtarget in a cell comprising providing to the cell a polypeptide, acomposition, a nucleic acid molecule, and/or the vector.

In another aspect, provided herein is a method of affecting translationof an RNA target in a cell comprising providing to a polypeptide, acomposition, a nucleic acid molecule, and/or a vector.

In some embodiments, the RNA target is an ssRNA.

In another aspect, provided herein is a method of treating a geneticallyinherited disease in a subject in need thereof comprising administeringto the subject an effective amount of a polypeptide, a composition, anucleic acid molecule, and/or a vector, wherein the geneticallyinherited disease involves a guanosine to adenosine change in a genomeof the subject.

In some embodiments, the genetically inherited disease is selected fromthe group consisting of Meier-Gorlin syndrome; Seckel syndrome 4;Joubert syndrome 5; Leber congenital amaurosis 10; Charcot-Marie-Toothdisease, type 2; leukoencephalopathy; Usher syndrome, type 2C;spinocerebellar ataxia 28; glycogen storage disease type III; primaryhyperoxaluria, type I; long QT syndrome 2; Sjögren-Larsson syndrome;hereditary fructosuria; neuroblastoma; amyotrophic lateral sclerosistype 9; Kallmann syndrome 1; limb-girdle muscular dystrophy, type 2L;familial adenomatous polyposis 1; familial type 3 hyperlipoproteinemia;Alzheimer's disease, type 1; metachromatic leukodystrophy; cancer;Uveitis; SCA1; SCA2; FUS-Amyotrophic Lateral Sclerosis (ALS);MAPT-Frontotemporal Dementia (FTD); Myotonic Dystrophy Type 1 (DM1);Diabetic Retinopathy (DR/DME); Oculopharyngeal Muscular Dystrophy(OPMD); SCA8; C9ORF72-Amyotrophic Lateral Sclerosis (ALS);SOD1-Amyotrophic Lateral Sclerosis (ALS); Spinal Cord Injury (targets:mTOR, PTEN, KLF6/7, SOX11, KCC2, and growth factors); SCA6; SCA3(Machado-Joseph Disease); Multiple system Atrophy (MSA);Treatment-resistant Hypertension; Myotonic Dystrophy Type 2 (DM2);Fragile X-associated Tremor Ataxia Syndrome (FXTAS); West Syndrome withARX Mutation; Age-related Macular Degeneration (AMD)/Geographic Atrophy(GA); C9ORF72-Frontotemporal Dementia (FTD); FacioscapulohuneralMuscular Dystrophy (FSHD); Fragile X Syndrome (FXS); Huntington'sDisease; Glaucoma; Acromegaly; Achromatopsia (total color blindness);Ullrich congenital muscular dystrophy; Hereditary myopathy with lacticacidosis; X-linked spondyloepiphyseal dysplasia tarda; Neuropathic pain(Target: CPEB); Persistent Inflammation and injury pain (Target: PABP);Neuropathic pain (Target: miR-30c-5p); Neuropathic pain (Target:miR-195); Friedreich's Ataxia; Uncontrolled gout; Inflammatory pain(Target: Nav1.7 and Nav1.8); Choroideremia; Focal epilepsy; Alpha-1Antitrypsin deficiency (AATD); Androgen Insensitivity Syndrome;Opioid-induced hyperalgesia (Target: Raf-1); Neurofibromatosis type 1;Stargardt's Disease; Dravet Syndrome; Retinitis Pigmentosa; andParkinson's Disease.

In another aspect, provided herein is a method of treating a geneticallyinherited disease in a subject in need thereof comprising administeringto the subject an effective amount of a polypeptide, a composition, anucleic acid molecule, and/or a vector, wherein the geneticallyinherited disease is a pre-termination disease.

In another aspect, provided herein is a method of altering splicing of apre-mRNA in a cell comprising administering to the cell an effectiveamount of a polypeptide, a composition, a nucleic acid molecule, and/ora vector.

In another aspect, provided herein is a method of changing microRNAtargets in a subject in need thereof comprising administering to thesubject an effective amount of a polypeptide, a composition, a nucleicacid molecule, and/or a vector.

In another aspect, provided herein is a method of increasing RNAstability in a cell comprising administering to the cell an effectiveamount of a polypeptide, a composition, a nucleic acid molecule, and/ora vector.

In another aspect, provided herein is a method of modulating translationin a cell comprising administering to the cell an effective amount of apolypeptide, a composition of, a nucleic acid molecule, and/or a vector.

In another aspect, provided herein is a method of detecting a bacteriumor derivative thereof in a sample, the method comprising: adding to thesample an effective amount of a polypeptide, a composition, a nucleicacid molecule, and/or a vector; and detecting a reporter specific to thebacterium or derivative thereof.

In another aspect, provided herein is a method of detecting a virus orderivative thereof in a sample, the method comprising: adding to thesample an effective amount of a polypeptide, a composition, a nucleicacid molecule, and/or a vector; and detecting a reporter specific to thevirus or derivative thereof.

These and other aspects of the applicants' teaching are set forthherein.

BRIEF DESCRIPTION OF THE DRAWINGS

Aspects, features, benefits and advantages of the embodiments describedherein will be apparent with regard to the following description,appended claims, and accompanying drawings where:

FIG. 1A is a schematic diagram of a domain structure of Cas7-11;

FIG. 1B is a schematic diagram of nucleotide sequences of a crRNA (SEQID NO: 8) and its target RNA (SEQ ID NO: 5), wherein a pre-crRNAprocessing site and target RNA cleavage sites are indicated by cyan, andyellow and green triangles, respectively, and a crRNA and target RNAcontain a 5′ GG for in vitro transcription;

FIG. 1C a ribbon representation of an overall structure of aCas7-11-crRNA-target RNA complex, wherein zinc ions bound to theCas7.1-Cas7.4 domains are shown as orange spheres, the disorderedregions are indicated as dotted lines, and the disordered L1 and L2linkers are not shown for clarity;

FIG. 1D is a surface representation of a Cas7-11-crRNA-target RNAcomplex;

FIG. 2A is a ribbon representation of a Cas7.1 domain;

FIG. 2B is a ribbon representation of a Cas7.2 domain;

FIG. 2C is a ribbon representation of a Cas7.3 domain;

FIG. 2D is a ribbon representation of a Cas7.4 domain;

FIG. 2E is a ribbon representation of a Cas11 domain;

FIG. 2F is a ribbon representation of an INS domain;

FIG. 3A is a schematic representation of crRNA and target RNAstructures;

FIG. 3B is a schematic representation of interactions between Cas7-11and bound nucleic acids, wherein Cas7-11 residues that interact withnucleic acids through their main chains are shown in parentheses, and apre-crRNA processing site and target RNA cleavage sites are indicated bycyan, and yellow and green triangles, respectively;

FIG. 4A is a surface representation of a crRNA 5′ tag region, wherein apre-crRNA processing site is indicated by a cyan triangle, and pGp isguanosine-3′,5′-diphosphate;

FIG. 4B is a ribbon representation of a C(−1)-G(−4) region in a crRNA 5′tag;

FIG. 4C is a ribbon representation of a C(−6) region in a crRNA 5′ tag;

FIG. 4D is a ribbon representation of a A(−7)-U(−9) region in a crRNA 5′tag;

FIG. 4E is a ribbon representation of a G(−10)-U(−14) region in a crRNA5′ tag;

FIG. 4F is a ribbon representation of a pGp molecule, wherein densitiesfor the pGp and crRNA molecules are shown as gray meshes, and apre-crRNA processing site is indicated by a cyan triangle;

FIG. 4G is a schematic representation and an image of an electrophoresisgel for an in vitro pre-crRNA processing by WT Cas7-11 and Cas7-11mutants, wherein target RNA cleavage activities were measured using amature crRNA containing a 14-nt 5′ tag with a 5′ GG, or a pre-crRNAcontaining a 23-nt 5′ tag with a 5′ GG TBC;

FIG. 4H is a schematic representation and an image of an electrophoresisgel for an in vitro target RNA cleavage by WT Cas7-11 and Cas7-11mutants;

FIG. 5A is a surface representation of a guide-target duplex byCas7.4/INS/CTE domains, wherein target RNA cleavage sites (sites 1 and2) are indicated by yellow and green triangles;

FIG. 5B is a ribbon representation of a guide-target duplex byCas7.4/INS/CTE domains, wherein target RNA cleavage sites (sites 1 and2) are indicated by yellow and green triangles;

FIG. 5C is a surface representation of a guide-target duplex byCas7.2-Cas7.4/Cas11 domains, wherein target RNA cleavage sites (sites 1and 2) are indicated by yellow and green triangles;

FIG. 5D is a ribbon representation of a guide-target duplex byCas7.2-Cas7.4/Cas11 domains, wherein target RNA cleavage sites (sites 1and 2) are indicated by yellow and green triangles;

FIG. 5E is a schematic representation and an image of an electrophoresisgel for an in vitro target RNA cleavage by a WT Cas7-11 usingmismatch-containing crRNAs;

FIG. 6A are ribbon representations of Cas7-11 variants;

FIG. 6B are images of an electrophoresis gel for an in vitro target RNAcleavage by truncated Cas7-11 variants;

FIG. 6C is a diagram showing the knockdown of Gluc mRNA by truncatedCas7-11 variants in HEK293FT cells, wherein data are normalized tonon-targeting controls and shown as mean±s.e.m. (n=3);

FIG. 6D is a diagram showing the knockdown of endogenous mRNAtranscripts by truncated Cas7-11 variants in HEK293FT cells, whereindata are normalized to non-targeting controls and shown as mean±s.e.m.(n=3);

FIG. 6E are schematic representations of an AAV design;

FIG. 6F is a schematic representation an AAV experimental;

FIG. 6G is a diagram showing the knockdown of Gluc mRNA by truncatedCas7-11 variants in HEK293FT cells via AAV delivery, wherein data arenormalized to non-targeting controls and shown as mean±s.e.m. (n=3);

FIG. 7A are schematic representations of a structure of a type III-ECas7-11 complex, wherein guide-target duplexes are shown on the left ofthe complexes;

FIG. 7B are schematic representations of a structure of a type III-A Csmcomplex, wherein guide-target duplexes are shown on the left of thecomplexes;

FIG. 8A is a schematic representation of a single-particle cryo-EM imageprocessing workflow;

FIG. 8B is a graph showing a Fourier shell correlation curve calculatedbetween half-maps in a 3D reconstruction;

FIG. 8C is a graph showing a Fourier shell correlation curve calculatedbetween a refined model and a density map;

FIG. 8D is a surface representation of a local resolution of a densitymap;

FIG. 9A are ribbon representations of Cas7.2/Cas7.3 domains of Cas7-11and Csm3 subunits of Csm complex (PDB ID: 6IFY);

FIG. 9B are ribbon representations of Cas11 domain of Cas7-11 and Csm2subunit of Csm complex (PDB ID: 6IFY);

FIG. 9C are ribbon representations of INS domain of Cas7-11 and Bacillussubtilis cold shock protein (CSP) (PDB ID: 1CSP), wherein the INS domaincontains the two five-stranded β-barrels similar to cold shock proteins;

FIG. 10A (SEQ ID NO:110, 111, 112, 113) shows multiple sequencealignments of Cas7-11 orthologs wherein the alignments were preparedusing Clustal Omega (http://www.ebi.ac.uk/Tools/msa/clustalo) andESPript3 (http://espript.ibcp.fr/ESPript/ESPript), and key residues aremarked triangles,

FIG. 10B (SEQ ID NO:110, 111, 112, 113) shows multiple sequencealignments of Cas7-11 orthologs wherein the alignments were preparedusing Clustal Omega (http://www.ebi.ac.uk/Tools/msa/clustalo) andESPript3 (http://espript.ibcp.fr/ESPript/ESPript), and key residues aremarked triangles;

FIG. 10C (SEQ ID NO:110, 111, 112, 113) shows multiple sequencealignments of Cas7-11 orthologs wherein the alignments were preparedusing Clustal Omega (http://www.ebi.ac.uk/Tools/msa/clustalo) andESPript3 (http://espript.ibcp.fr/ESPript/ESPript), and key residues aremarked triangles;

FIG. 11A is a surface representation of Cas7.1/Cas7.3 domain, whereinthe crRNA and target RNA are omitted for clarity;

FIG. 11B is a surface representation of Cas7.2/Cas7.4/INS domain,wherein the crRNA and target RNA are omitted for clarity;

FIG. 11C is a surface representation of Cas11/CTE domain, wherein thecrRNA and target RNA are omitted for clarity;

FIG. 11D is a surface representation of Cas11/L4 domain, wherein thecrRNA and target RNA are omitted for clarity;

FIG. 11E is a schematic representation of Cas7-11 domains;

FIG. 12 is a density map (unsharpened, FSC-weighted) for bound RNAmolecules (stereo view);

FIG. 13A is a ribbon representation of a guide-target duplex by theCas7.2/Cas7.3/Cas11 domains of type III-E Cas7-11, wherein target RNAcleavage sites (sites 1 and 2) are indicated by yellow and greentriangles, respectively;

FIG. 13B is a ribbon representation of a guide-target duplex by theCsm1/Csm2/Csm3 subunits of type III-A Csm (stereoview), wherein targetRNA cleavage sites (sites 1 and 2) are indicated by yellow and greentriangles, respectively;

FIG. 13C shows the superposition of the Cas7-11 and Csm complexes(stereo view);

FIG. 14 shows a schematic representation and an image of anelectrophoresis gel for the pr-crRNA processing by DiCas7-11 mutants;

FIG. 15 shows a schematic representation and an image of anelectrophoresis gel for the target ssRNA cleavage by pre-mature andmature crRNA with DiCas7-11 mutants;

FIG. 16 shows a schematic representation and images of electrophoresisgels for the target ssRNA cleavage by mismatched crRNA guides andDiCas7-11;

FIG. 17 shows images of electrophoresis gels for the target ssRNAcleavage by mismatched crRNA guides and DiCas7-11 mutants;

FIG. 18A shows images of electrophoresis gels for the target ssRNAcleavage by mismatched crRNA guides and mismatched target;

FIG. 18B shows images of electrophoresis gels for the target ssRNAcleavage by mismatched crRNA guides and mismatched target;

FIG. 19A is a schematic representation of the target ssRNA cleavage byDiCas7-11 processing mutants and truncated DiCas7-11;

FIG. 19B shows images of electrophoresis gels for the target ssRNAcleavage by DiCas7-11 processing mutants and truncated DiCas7-11;

FIG. 20A shows a schematic representation of the knockdown of Gluc mRNAin HEK293FT cells by truncated DiCas7-11;

FIG. 20B shows a diagram for the knockdown of Gluc mRNA in HEK293FTcells by truncated DiCas7-11;

FIG. 21A shows a diagram for the knockdown of endogenous mRNA inHEK293FT cells by truncated DiCas7-11 for PPIB transcript;

FIG. 21B shows a diagram for the knockdown of endogenous mRNA inHEK293FT cells by truncated DiCas7-11 for MALAT1 transcript;

FIG. 21C shows a diagram for the knockdown of endogenous mRNA inHEK293FT cells by truncated DiCas7-11 for transcript;

FIG. 22A shows A diagram for the knockdown of Gluc mRNA in HEK293FTcells by truncated DiCas7-11 packaged in AAV8 vector;

FIG. 22B shows a diagram for the knockdown of Gluc mRNA in HEK293FTcells by truncated DiCas7-11 packaged in AAV8 vector;

FIG. 23 shows a schematic representation and an intensity graph for theknockdown of Gluc mRNA in HEK293FT cells by DiCas7-11 and mismatchedcrRNA guides;

FIG. 24 shows a ribbon representation of DiCas7-11 and the location ofresidue mutations;

FIG. 25 shows a diagram of single mutant g-luciferase knockdown;

FIG. 26 shows a diagram of single mutant endogenous MALAT1 knockdown;

FIG. 27 shows a diagram of single mutant endogenous PPIB knockdown;

FIG. 28 shows a diagram of single and double mutant G-luciferaseknockdown;

FIG. 29 shows a diagram of saturation mutagenesis for D1530 residue; and

FIG. 30 shows a diagram of single, double, triple, and quadruple mutantsG-luciferase knockdown.

DETAILED DESCRIPTION

It will be appreciated that for clarity, the following discussion willdescribe various aspects of embodiments of the applicant's teachings. Itshould be noted that the specific embodiments are not intended as anexhaustive description or as a limitation to the broader aspectsdiscussed herein. One aspect described in conjunction with a particularembodiment is not necessarily limited to that embodiment and can bepracticed with any other embodiment(s).

General Definitions

Unless defined otherwise, technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure pertains. Definitions of common termsand techniques in molecular biology may be found in Molecular Cloning: ALaboratory Manual, 2^(nd) edition (1989) (Sambrook, Fritsch, andManiatis); Molecular Cloning: A Laboratory Manual, 4^(th) edition (2012)(Green and Sambrook); Current Protocols in Molecular Biology (1987) (F.M. Ausubel et al. eds.); the series Methods in Enzymology (AcademicPress, Inc.): PCR 2: A Practical Approach (1995) (M. J. MacPherson, B.D. Hames, and G. R. Taylor eds.): Antibodies, A Laboratory Manual (1988)(Harlow and Lane, eds.): Antibodies A Laboratory Manual, 2^(nd) edition2013 (E. A. Greenfield ed.); Animal Cell Culture (1987) (R. I. Freshney,ed.); Benjamin Lewin, Genes I X, published by Jones and Bartlet, 2008(ISBN 0763752223); Kendrew et al. (eds.), The Encyclopedia of MolecularBiology, published by Blackwell Science Ltd., 1994 (ISBN 0632021829);Robert A. Meyers (ed.), Molecular Biology and Biotechnology: aComprehensive Desk Reference, published by VCH Publishers, Inc., 1995(ISBN 9780471185710); Singleton et al., Dictionary of Microbiology andMolecular Biology 2^(nd) ed., J. Wiley & Sons (New York, N.Y. 1994),March, Advanced Organic Chemistry Reactions, Mechanisms and Structure4^(th) ed., John Wiley & Sons (New York, N.Y. 1992); and Marten H.Hofker and Jan van Deursen, Transgenic Mouse Methods and Protocols,2^(nd) edition (2011).

As used herein, the singular forms “a”, “an,” and “the” include bothsingular and plural referents unless the context clearly dictatesotherwise.

The term “optional” or “optionally” means that the subsequent describedevent, circumstance or substituent may or may not occur, and that thedescription includes instances where the event or circumstance occursand instances where it does not.

The recitation of numerical ranges by endpoints includes all numbers andfractions subsumed within the respective ranges, as well as the recitedendpoints.

The terms “about” or “approximately” as used herein when referring to ameasurable value such as a parameter, an amount, a temporal duration,and the like, are meant to encompass variations of and from thespecified value, such as variations of +/−10% or less, +1-5% or less,+/−1% or less, +1-0.5% or less, and +/−0.1% or less of and from thespecified value, insofar such variations are appropriate to perform inthe disclosed disclosure. It is to be understood that the value to whichthe modifier “about” or “approximately” refers is itself alsospecifically, and preferably, disclosed.

Overview

The embodiments disclosed herein provide (non-naturally occurring orengineered) constructs, compositions, systems, and methods forsite-directed RNA editing of RNA molecules. For example, the presentdisclosure provides (non-naturally occurring or engineered) methods forinhibiting intra and inter-cellular signaling pathways by modificationof post-translational modification sites on select target RNA molecules.In certain embodiments, the present disclosure provides (non-naturallyoccurring or engineered) methods for inhibiting intracellularphosphorylation of serine, threonine and tyrosine residues by editingthe genetic codon of these amino acids by means of site-directed RNAediting or RNA molecules. Embodiments disclosed herein further providemethods of inhibiting pathological activation of cell signaling mediatedby post-translational modifications, such as phosphorylation, which areinvolved in many diseases, including cancer, immunodeficiency,infectious diseases, inflammatory disorders and neurodegenerativedisorders. The RNA-editing modification may be aimed at a singlepost-translational modification site of a single gene and can also bemultiplexed by targeting multiple sites on the same or different genesto increase efficacy. These approaches may be further combined withother treatments such as radiation, chemotherapy, targeted therapy basedon antibodies or small molecules, and immunotherapy, which may have asynergistic effect.

The embodiments disclosed herein provide (non-naturally occurring orengineered) systems, constructs, and methods for targeted base editing.In general, the systems disclosed herein comprise a targeting componentand a base editing component. The targeting component may function tospecifically target the base editing component to a target nucleotidesequence in which one or more nucleotides are to be edited. The baseediting component may then catalyze a chemical reaction to convert afirst nucleotide in the target sequence to a second nucleotide. Forexample, the base editor may catalyze conversion of an adenine such thatit is read as guanine by a cell's transcription or translationmachinery, or vice versa. Likewise, the base editing component maycatalyze conversion of cytidine to an uracil, or vice versa. In certainexample embodiments, the base editor may be derived by starting with aknown base editor, such as an adenine deaminase or cytidine deaminase,and modified using methods such as directed evolution to derive newfunctionalities. Directed evolution techniques are known in the art andmay include those described in WO 2015/184016 “High-Throughput Assemblyof Genetic Permutations.”

Compositions and Systems

The present disclosure provides (non-naturally occurring or engineered)systems for editing a nucleic acid such as a gene or a product thereof(e.g., the encoded RNA or protein).

In some embodiments, the systems may be an engineered, non-naturallyoccurring system suitable for modifying post-translational modificationsites on proteins encoded by a target nucleic acid sequence. In certaincases, the target nucleic acid sequence is RNA, e.g., mRNA or a fragmentthereof. In certain cases, the target nucleic acid sequence is DNA,e.g., a gene or a fragment thereof. In general, the system may compriseone or more of a catalytic inactive (dead) Cas protein (e.g., deadCas7-11), a nucleotide deaminase protein or catalytic domain thereof,and a guide molecule. In certain examples, the nucleotide deaminaseprotein may be an adenosine deaminase. In certain examples, thenucleotide deaminase protein may be a cytidine deaminase. The guidesequence may be designed to have a degree of complementarity with atarget sequence at one or more codons comprising an adenine or cytidineand that is post-translationally modified.

CRISPR-Cas

Some embodiments disclosed herein are directed to CRISPR-Cas (clusteredregularly interspaced short palindromic repeats associated proteins)systems. In the conflict between bacterial hosts and their associatedviruses, CRISPR-Cas systems provide an adaptive defense mechanism thatutilizes programmed immune memory. CRISPR-Cas systems provide theirdefense through three stages: adaptation, the integration of shortnucleic acid sequences into the CRISPR array that serves as memory ofpast infections; expression, the transcription of the CRISPR array intoa pre-crRNA (CRISPR RNA) transcript and processing of the pre-crRNA intofunctional crRNA species targeting foreign nucleic acids; andinterference, the programming of CRISPR effectors by crRNA to cleavenucleic acid of foreign threats. Across all CRISPR-Cas systems, thesefundamental stages display enormous variation, including the identity ofthe target nucleic acid (either RNA, DNA, or both) and the diversedomains and proteins involved in the effector ribonucleoprotein complexof the system.

CRISPR-Cas systems can be broadly split into two classes based on thearchitecture of the effector modules involved in pre-crRNA processingand interference. Class 1 systems have multi-subunit effector complexescomposed of many proteins, whereas Class 2 systems rely onsingle-effector proteins with multi-domain capabilities for crRNAbinding and interference; Class 2 effectors often provide pre-crRNAprocessing activity as well. Class 1 systems contain 3 types (type I,III, and IV) and 33 subtypes, including the RNA and DNA targeting typeIII-systems. Class 2 CRISPR families encompass 3 types (type II, V, andVI) and 17 subtypes of systems, including the RNA-guided Dnases Cas9 andCas12 and the RNA-guided Rnase Cas13. Continual sequencing of novelbacterial genomes and metagenomes uncovers new diversity of CRISPR-Cassystems and their evolutionary relationships, necessitating experimentalwork that reveals the function of these systems and develops them intonew tools.

Among the currently known CRISPR-Cas systems, only the type III and typeVI systems have been demonstrated to bind and target RNA, and these twosystems have substantially different properties, the most distinguishingbeing their membership in Class 1 and Class 2, respectively.Characterized subtypes of type III, which span type III-A, B, and Csystems, target both RNA and DNA species through an effector complexcontaining multiple Cas7 (Csm3/5 or Cmr1/4/6) RNA nuclease units inassociation with a single Cas10 (Csm1 or Cmr2) DNA nuclease. The RNAnuclease activity of Cas7 is mediated through acidic residues in therepeat-associated mysterious proteins (RAMP) domains, which cut atstereotyped intervals in the guide:target duplex. Type III systems alsohave a target restriction and cannot efficiently target protospacers invivo if there is extended homology between the 5′ “tag” of the crRNA andthe “anti-tag” 3′ of the protospacer in the target, although thisbinding does not block RNA cleavage in vitro. In type III systems,pre-crRNA processing is carried out by either host factors or theassociated Cas6 family protein, which can physically complex with theeffector machinery.

In contrast to type III systems, type VI systems contain a single CRISPReffector Cas13 that can only effect RNA interference, mediated throughbasic catalytic residues of dual HEPN domains. This interferencerequires a protospacer flanking sequence (PFS), although the influenceof the PFS varies between orthologs and families. Importantly, the RNAcleavage activity of Cas13, once triggered by crRNA:target duplexformation, is indiscriminate, and activated Cas13 enzymes will cleaveother RNA species in vitro, in bacterial hosts, and mammalian cells.This activity, termed the collateral effect, has been applied toCRISPR-based nucleic acid detection technologies. In addition to the RNAinterference activity, the Cas13 family members contain pre-crRNAprocessing activity. Just as single-effector DNA targeting systems havegiven rise to numerous genome editing applications, Cas13 family membershave been applied to a suite of RNA-targeting technologies in bothbacterial and eukaryotic cells, including RNA knockdown, RNA editing,RNA tracking, epitranscriptome editing, translational upregulation,epi-transcriptomic reading and writing via N6-Methyladenosine, andisoform modulation.

The novel type III-E system was recently identified from genomes of 8bacterial species and is characterized as a fusion of several Cas7proteins and a putative Cas11 (Csm2)-like small subunit. The domaincomposition suggests the fusion of multiple type III effector moduledomains involved in crRNA binding into a single protein effector that ispredicted to process pre-crRNA given its homology with Cas5 (Csm4) andconserved aspartates. The lack of other putative effector nucleases inthese CRISPR loci raise the additional possibility that this fusionprotein is capable of crRNA-directed RNA cleavage. If so, this systemwould blur the distinction of Class 1 and Class 2 systems, as it wouldhave domains homologous to other Class 1 systems and possess a singleeffector module characteristic of Class 2 systems. Beyond the singleeffector module present in all subtype III-E loci, a majority of typeIII-E family members contain a putative ancillary gene with a CHATdomain, which is a caspase family protease associated with programmedcell death (PCD), suggesting involvement of PCD-mediated antiviralstrategies, as has been observed with type III and VI systems.

Type III-E system associated effector is a programmable Rnase. Thissystem can provide defense against RNA phage and be programmed to targetexogenous mRNA species when expressed heterologously in bacteria.Orthologs of Cas7-11 are capable of both processing of pre-crRNA andcrRNA-directed cleavage of RNA targets and determine catalytic residuesunderlying programmed RNA cleavage. A direct evolutionary path ofCas7-11 can be traced from individual Cas7 and Cas11 effector proteinsof subtype III-D1 variant, through an intermediate, a partially fusedeffector Cas7×3 of the subtype III-D2 variant, to the singe-effectorarchitecture of subtype III-E that is so far unique among the Class 1CRISPR-Cas systems. Cas7-11 most likely originated from two type III-Dvariants. Three Cas7 domains (domains 3, 4 and 5) are derived fromsubtype III-D2 that contains a the Cas7×3 effector protein along withCas10 and another Cas7-like domain fused to a Cas5-like domain. Theorigin of the N-terminal Cas7 and putative Cas11 domain of Cas7-11 ismost likely derived from a III-D1 variant, where both genes arestand-alone.

Cas7-11 differs from Cas13, in terms of both domain organization andactivity. Cas13 RNA cleavage is enacted by dual HEPN domains with basiccatalytic residues, and this cleavage, once triggered, isindiscriminate. In contrast, Cas7-11 utilizes at least two of fourCas7-like domains with acidic catalytic residues to generate stereotypedcleavage at the target binding site in cis. Furthermore, Cas13 targetingis restricted by the requirement for a PFS, which Cas7-11 does notrequire, and the DR of Cas7-11-associated crRNA is substantiallyshorter. Because of these unique features, Cas7-11 may have distinctadvantages for RNA targeting and transcriptome engineering biotechnologyapplications.

Regulation of interference by accessory proteins has been observed inboth type III and type VI systems, and other proteins in the D.ishimotonii type III-E locus can regulate activity of DiCas7-11a.Notably, TPR-CHAT had a strong inhibitory effect on DiCas7-11a phageinterference, raising the possibility that unrestricted DiCas7-11aactivity could be detrimental for the host. Alternatively, as TPR-CHATis a caspase family protease associated with programmed cell death(PCD), it is possible that TPR-CHAT is activated by DiCas7-11a and leadsto host death, which could mimic death due to phage in these assays.TPR-CHAT caspase activity could be activated by DiCas7-11a and cause PCDthrough general proteolysis, analogous to PCD triggered by Cas13collateral activity.

Similar to Class 2 CRISPR effectors such as Cas9, Cas12, and Cas13,Cas7-11 is highly active in mammalian cells, with substantial knockdownactivity on both reporter and endogenous transcripts. Moreover, viainactivation of active sites through mutagenesis, the catalyticallyinactive dCas7-11 enzyme can be used to recruit ADAR2DD for efficientsite-specific A-to-I editing on transcripts. These applicationsestablish Cas7-11 as the basis for an RNA-targeting toolbox that hasseveral benefits compared to Cas13, including the lack of sequencepreferences and collateral activity, the latter of which has been shownto induce toxicity in certain cell types. A Cas7-11 toolbox may serve asthe basis for multiple RNA technologies, including RNA knockdown, RNAediting, translation modulation, RNA recruitment, RNA tracking, splicingcontrol, RNA stabilization, and potentially even diagnostics.

AD-Functionalized CRISPR Systems

In some embodiments, the systems may be AD-functionalized CRISPR system.The term “AD-functionalized CRISPR system” as used here refers to anucleic acid targeting and editing system comprising (a) a CRISPR-Casprotein, more particularly a Cas7-11 protein which is catalyticallyactive or inactive; (b) a guide molecule which comprises a guidesequence; and (c) an adenosine deaminase (AD) protein or catalyticdomain thereof; wherein the adenosine deaminase protein or catalyticdomain thereof is covalently or non-covalently linked to the CRISPR-Casprotein or the guide molecule or is adapted to link thereto afterdelivery; wherein the guide sequence is substantially complementary tothe target sequence but comprises a non-pairing C corresponding to the Abeing targeted for deamination, resulting in an A-C mismatch in an RNAduplex formed by the guide sequence and the target sequence. In someembodiments, the CRISPR-Cas protein and/or the adenosine deaminasecomprise one or more heterologous nuclear export signal(s) (NES(s)) ornuclear localization signal(s)(NLS(s)). For application in eukaryoticcells, the CRISPR-Cas protein and/or the adenosine deaminase can beNES-tagged or NLS-tagged.

One skilled in the art would appreciate that the components (a), (b) and(c) can be delivered to the cell as a ribonucleoprotein complex. Theribonucleoprotein complex can be delivered via one or more lipidnanoparticles. One skilled in the art would appreciate that thecomponents (a), (b) and (c) can be delivered to the cell as one or moreRNA molecules, such as one or more guide RNAs and one or more mRNAmolecules encoding the CRISPR-Cas protein, the adenosine deaminaseprotein, and optionally the adaptor protein. The RNA molecules can bedelivered via one or more lipid nanoparticles. One skilled in the artwould appreciate that the components (a), (b) and (c) can be deliveredto the cell as one or more DNA molecules. The one or more DNA moleculescan be comprised within one or more vectors such as viral vectors (e.g.,AAV). The one or more DNA molecules can comprise one or more regulatoryelements operably configured to express the CRISPR-Cas protein, theguide molecule, and the adenosine deaminase protein or catalytic domainthereof, optionally wherein the one or more regulatory elements compriseinducible promoters.

In some embodiments, the CRISPR-Cas protein is a dead Cas7-11. In someembodiments, the dead Cas7-11 comprises one or more mutations in theCas7-like domains, including D429A and D654A as well as many othermutations.

In some embodiments, the CRISPR-Cas protein is a Cas7-11 endonucleasewith an amino acid sequence comprising at least 1 mutation ormodification, at least 2 mutations or modifications, at least 3mutations or modifications, at least 4 mutations or modifications, atleast 5 mutations or modifications, at least 6 mutations ormodifications, at least 7 mutations or modifications, at least 8mutations or modifications, at least 9 mutations or modifications, atleast 10 mutations or modifications, or any ranges that are made of anytwo or more points in the above list of mutations or modifications. Insome embodiments, the Cas7-11 endonuclease is a DiCas7-11 endonuclease.

In some embodiments, the guide molecule is capable of hybridizing with atarget sequence comprising the Adenine to be deaminated within an RNAsequence to form an RNA duplex which comprises a non-pairing Cytosineopposite to said Adenine. Upon RNA duplex formation, the guide moleculeforms a complex with the Cas7-11 protein and directs the complex to bindthe RNA polynucleotide at the target RNA sequence of interest. Detailson the aspect of the guide of the AD-functionalized CRISPR-Cas systemare provided herein below.

In at least a first design, the AD-functionalized CRISPR systemcomprises: (a) an adenosine deaminase fused or linked to a CRISPR-Casprotein, wherein the CRISPR-Cas protein is catalytically inactive; and(b) a guide molecule comprising a guide sequence designed to introducean A-C mismatch in an RNA duplex formed between the guide sequence andthe target sequence. In some embodiments, the CRISPR-Cas protein and/orthe adenosine deaminase can be NLS-tagged on either the N- or C-terminusor both.

In at least a second design, the AD-functionalized CRISPR systemcomprises: (a) a CRISPR-Cas protein that is catalytically inactive; (b)a guide molecule comprising a guide sequence designed to introduce anA-C mismatch in an RNA duplex formed between the guide sequence and thetarget sequence, and an aptamer sequence (e.g., MS2 RNA motif or PP7 RNAmotif) capable of binding to an adaptor protein (e.g., MS2 coatingprotein or PP7 coat protein); and (c) an adenosine deaminase fused orlinked to an adaptor protein, wherein the binding of the aptamer and theadaptor protein recruits the adenosine deaminase to the RNA duplexformed between the guide sequence and the target sequence for targeteddeamination at the A of the A-C mismatch. In some embodiments, theadaptor protein and/or the adenosine deaminase can be NLS-tagged oneither the N- or C-terminus or both. The CRISPR-Cas protein can also beNLS-tagged. The CRISPR-Cas protein can also be NLS-tagged.

The use of different aptamers and corresponding adaptor proteins alsoallows orthogonal gene editing to be implemented. In one example inwhich adenosine deaminase are used in combination with cytidinedeaminase for orthogonal gene editing/deamination, sgRNA targetingdifferent loci are modified with distinct RNA loops in order to recruitMS2-adenosine deaminase and PP7-cytidine deaminase (or PP7-adenosinedeaminase and MS2-cytidine deaminase), respectively, resulting inorthogonal deamination of A or C at the target loci of interested,respectively. PP7 is the RNA-binding coat protein of the bacteriophagePseudomonas. Like MS2, it binds a specific RNA sequence and secondarystructure. The PP7 RNA-recognition motif is distinct from that of MS2.Consequently, PP7 and MS2 can be multiplexed to mediate distinct effectsat different genomic loci simultaneously. For example, an sgRNAtargeting locus A can be modified with MS2 loops, recruitingMS2-adenosine deaminase, while another sgRNA targeting locus B can bemodified with PP7 loops, recruiting PP7-cytidine deaminase. In the samecell, orthogonal, locus-specific modifications are thus realized. Thisprinciple can be extended to incorporate other orthogonal RNA-bindingproteins.

In at least a third design, the AD-functionalized CRISPR systemcomprises: (a) an adenosine deaminase inserted into an internal loop orunstructured region of a CRISPR-Cas protein, wherein the CRISPR-Casprotein is catalytically inactive or a nickase; and (b) a guide moleculecomprising a guide sequence designed to introduce an A-C mismatch in anRNA duplex formed between the guide sequence and the target sequence.

The AD-functionalized CRISPR system described herein can be used totarget a specific Adenine within an RNA polynucleotide sequence fordeamination. For example, the guide molecule can form a complex with theCRISPR-Cas protein and directs the complex to bind a target RNA sequencein the RNA polynucleotide of interest. Because the guide sequence isdesigned to have a non-pairing C, the RNA duplex formed between theguide sequence and the target sequence comprises an A-C mismatch, whichdirects the adenosine deaminase to contact and deaminate the A oppositeto the non-pairing C, converting it to an Inosine (I). Since Inosine (I)base pairs with C and functions like G in cellular processes, thetargeted deamination of A described herein are useful for correction ofundesirable G-A and C-T mutations, as well as for obtaining desirableA-G and T-C mutations.

In some embodiments, the AD-functionalized CRISPR system is used fortargeted deamination in an RNA polynucleotide molecule in vitro. In someembodiments, the AD-functionalized CRISPR system is used for targeteddeamination in a DNA molecule and/or RNA molecule within a cell. Thecell can be a eukaryotic cell such as a bacteria or cyanobacteria. Thecell can be a eukaryotic cell, such as an animal cell, a mammalian cell,a human, or a plant cell.

The disclosure also relates to a (non-naturally occurring or engineered)method for treating or preventing a disease by the targeted deaminationusing the AD-functionalized CRISPR system, wherein the deamination ofthe A, which remedies a disease caused by transcripts containing apathogenic G→A or C→T point mutation. Examples of disease that can betreated or prevented with the present disclosure include cancer,Meier-Gorlin syndrome, Seckel syndrome 4, Joubert syndrome 5, Lebercongenital amaurosis 10; Charcot-Marie-Tooth disease, type 2;Charcot-Marie-Tooth disease, type 2; Usher syndrome, type 2C;Spinocerebellar ataxia 28; Spinocerebellar ataxia 28; Spinocerebellarataxia 28; Long QT syndrome 2; Sjogren-Larsson syndrome; Hereditaryfructosuria; Hereditary fructosuria; Neuroblastoma; Neuroblastoma;Kallmann syndrome 1; Kallmann syndrome 1; Kallmann syndrome 1;Metachromatic leukodystrophy.

AD-functionalized CRISPR system for RNA editing can be used fortranslation upregulation or downregulation, improving RNA stability anddiagnostics. For example, for application in diagnostics, TPR-Chat is anaccessory protein that interacts with Cas7-11 interference. Cas7-11 canactivate TPR-Chat caspase activity which can then activate a reporter.While this can be used for inducing cell death based on RNA detection(e.g., in cancer cells), it also can be useful for general RNAdiagnostics (i.e., molecular diagnostics for bacteria, viruses, andderivatives thereof) in samples. Furthermore, Cas7-11 can re-constitutea split protein like GFP on a specific transcript.

AD-functionalized CRISPR system for RNA editing can be used to treat orprevent premature termination diseases. Pre-termination diseases arecharacterized by mutations in early stop codons, either through singlenucleotide polymorphisms that introduce termination, indels that changethe translational frame of the protein and generate new stop codons, oralternative splicing that preferentially introduces exons that haveearly termination. By removing stop codons generated in these ways via Ato I editing, RNA editing with ADAR could rescue diseases involvingpremature termination. In cases where SNPs are not G to A, but generatenonsense mutations, clinical benefit could be derived from changingnonsense mutations into missense mutations.

AD-functionalized CRISPR system for RNA editing can be used to changefertility mutations without germline editing. One advantage of RNAediting over DNA editing is in cases of SNPs affecting fertility, wherecorrection with genome editing would necessarily result in germlineediting, with potential ethical or safety implications. RNA editingcould correct these mutations without permanent effects on the genome,thereby circumventing these issues.

AD-functionalized CRISPR system for RNA editing can be used for splicingalteration. Pre-mRNA requires specific splice donor and acceptorsequences in order to undergo processing by the spliceosome. Spliceacceptor sites contain an invariant AG sequence that is necessary foracceptance of the attack by the splice donor sequence and intronremoval. By targeting Cas7-11-ADAR fusions to pre-mRNA and editing AGsplice acceptor sites to IG, it can be possible to inactivate the spliceacceptor site, resulting in skipping of the downstream exon. Thisapproach to splicing alteration has advantages over the current methodof exon skipping with chemically modified anti-sense oligos.Cas7-11-ADAR can be genetically encoded, allowing for long-term exonskipping. Additionally, Cas7-11-ADAR creates a mutation to promoteskipping, which can be more robust than masking of the splicedonor/acceptor site by a double stranded RNA, as is done with anti-senseoligos.

AD-functionalized CRISPR system for RNA editing can be used to alterneoantigens. Neoantigens in cancer are novel antigens that are expressedin tumor cells due to mutations that arise because of defective mismatchrepair. Engineering T cells against neoantigens is advantageous becausethe T cells will have no off-target activity and thus toxicity since theantigens are only expressed in the tumor cells. With RNA base editors,the Cas7-11-ADAR fusions can be targeted to cancer cells to introducemutations in transcripts that would introduce amino acid changes and newantigens that can be targeted using chimeric antigen receptor T cells.This approach is better than DNA base editors because it is transientand thus the risk of editing non-tumor cells permanently due tooff-target delivery is minimal.

AD-functionalized CRISPR system for RNA editing can be used to changemicroRNA targets for tumor suppressors. ADAR naturally edits mRNA togenerate or remove microRNA targets, thereby modulating expression.Programmable RNA editing can be used to up- or down-regulate microRNAtargets via altering of targeting regions. Additionally, microRNAsthemselves are natural substrates for ADAR, and programmable RNA editingof microRNAs can reduce or enhance the function on their correspondingtargets.

AD-functionalized CRISPR system for RNA editing can be used to makemultiple edits along a region. The Cas7-11-ADAR fusions can be preciselytargeted to edit specific adenosines by introducing a mismatch in theguide region across from the desired adenosine target and creating abubble that is favorable for A-to-I editing. By introducing multiple ofthese mismatches across different adenosine sites in the guide/targetduplex, it can be possible to introduce multiple mutations at once.

AD-functionalized CRISPR system for RNA editing can be used for thereversal of TAA (double A to G) for PTC. Many diseases that involvepretermination codon changes involve a TAA stop codon, which wouldrequire A-to-I changes to correct rather than the TAG or TGA stop codonswhich only need one A-to-I edit. Two approaches can be used to reversethe TAA stop codon. (1) As described in the previous section, twomismatches can be introduced in the guide against the two adenosines inthe TAA codon. (2) A two-guide array can be used to convert each of theadenosines to inosine sequentially. The first guide in the array cancontain a mutation against the first adenosine and the second guide canthen have complementarity to this change and have a mismatch against thesecond adenosine in the stop codon.

AD-functionalized CRISPR system for RNA editing can be used to treat orprevent cancer (GOF, LOF mutation reversal). Many oncogenic changes incancer involve G to A mutations that introduce gain of function or lossof function phenotypes to the mutated proteins. The RNA base editors arewell positioned to correct these changes and reduce oncogenesis.

RNA editing with ADAR can be used for the design of new basepreferences. Current ADAR1/2 proteins have been found to havesurrounding base preferences for catalytic activity, which may poseconstraints for certain applications. Rational mutagenesis or directedevolution of ADAR variants with altered or relaxed base preferences canincrease the versatility of programmable RNA editing.

AD-functionalized CRISPR system for RNA editing can comprise ADARmutants with increased activity in human cells. Although ADAR mutantswith altered activity in vitro or in yeast have been previouslyreported, screening or rational design of mutants with increasedactivity in the context of human cells can improve the efficiency orspecificity of ADAR-based programmable RNA editing constructs.

AD-functionalized CRISPR system for RNA editing can be used inbiological applications of inosine generation. The RNA editing with ADARgenerates inosine, which, when occurring multiple times in a transcript,can interact with endogenous biological pathways to increaseinflammation in cells and tissues. Generation of multiple inosine basescan increase inflammation, especially in cells where inflammation canlead to clearance. Additional inosine generation could also be used todestabilize transcripts.

AD-functionalized CRISPR system for RNA editing can be used in removingupstream start codons to promote protein expression of downstream ORF(ATG mutation). Anti-sense oligos have been used for blocking upstreamstart codon sites to promote protein expression at downstream startcodons. This allows the boosting of endogenous protein levels fortherapeutic purposes. Cas7-11-ADAR fusions could accomplish a similareffect by converting ATG sites to ITG (GTG) sites and thus removeupstream codons in endogenous transcripts and thus boost proteintranslation. So far, most therapeutic applications discussed have beenfor correcting G to A mutations or removing pre-termination sites. Thiswould be an application that allows for boosting gene expression. A goodexample is boosting fetal hemoglobin levels in sickle cell disease andthalassemias.

AD-functionalized CRISPR system for RNA editing can comprise themutagenesis of ADAR for C to U or any transition. It is possible throughrational mutagenesis or directed evolution that the ADARs listed in theortholog section could be made into C to U editors or editors of anybase transition.

In particular embodiments, the compositions described herein can be usedin therapy. This implies that the methods can be performed in vivo, exvivo or in vitro. In particular embodiments, the methods can be notmethods of treatment of the animal or human body or a method formodifying the germ line genetic identity of a human cell. In particularembodiments, when carrying out the method, the target RNA can be notcomprised within a human or animal cell. In particular embodiments, whenthe target is a human or animal target, the method can be carried out exvivo or in vitro.

CRISPR-Cas Proteins and Guides

In some embodiments, the system comprises one or more components of aCRISPR-Cas system. For example, the system may comprise a Cas protein, aguide molecule, or a combination thereof.

In the methods and systems of the present disclosure use is made of aCRISPR-Cas protein and corresponding guide molecule. More particularly,the CRISPR-Cas protein is a class 2 CRISPR-Cas protein. In certainembodiments, said CRISPR-Cas protein is a Cas7-11. The Cas7-11 may beCas7-11a, Cas7-11b, Cas7-11c, or Cas7-11d. The CRISPR-Cas system doesnot require the generation of customized proteins to target specificsequences but rather a single Cas protein can be programmed by guidemolecule to recognize a specific nucleic acid target, in other words theCas enzyme protein can be recruited to a specific nucleic acid targetlocus of interest using said guide molecule.

CRISPR-Cas Proteins

In some embodiments, the systems may comprise a CRISPR-Cas protein. Incertain examples, the CRISPR-Cas protein may be a catalytically inactive(dead) Cas protein. The catalytically inactive (dead) Cas protein mayhave impaired (e.g., reduced or no) nuclease activity. In some cases,the dead Cas protein may have nickase activity. In some cases, the deadCas protein may be dead Cas 15 protein. For example, the dead Cas 15 maybe dead Cas7-11a, dead Cas7-11b, dead Cas7-11c, or dead Cas7-11d. Insome embodiments, the system may comprise a nucleotide sequence encodingthe dead Cas protein.

In its unmodified form, a CRISPR-Cas protein is a catalytically activeprotein. This implies that upon formation of a nucleic acid-targetingcomplex (comprising a guide RNA hybridized to a target sequence) one orboth DNA strands in or near (e.g., within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,20, 50, or more base pairs from) the target sequence is modified (e.g.,cleaved). As used herein the term “sequence(s) associated with a targetlocus of interest” refers to sequences near the vicinity of the targetsequence (e.g., within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, or morebase pairs from the target sequence, wherein the target sequence iscomprised within a target locus of interest). The unmodifiedcatalytically active Cas7-11 protein generates a staggered cut, whereby“the cut sites are typically within the target sequence” Moreparticularly, the staggered cut is typically 13-23 nucleotides distal tothe PAM. In particular embodiments, the cut on the non-target strand is17 nucleotides downstream of the PAM (i.e. between nucleotide 17 and 18downstream of the PAM), while the cut on the target strand (i.e. strandhybridizing with the guide sequence) occurs a further 4 nucleotidesfurther from the sequence complementary to the PAM (this is 21nucleotides upstream of the complement of the PAM on the 3′ strand orbetween nucleotide 21 and 22 upstream of the complement of the PAM).

In the methods according to the present disclosure, the CRISPR-Casprotein is mutated with respect to a corresponding wild-type enzyme suchthat the mutated CRISPR-Cas protein lacks the ability to cleave one orboth DNA strands of a target locus containing a target sequence. Inparticular embodiments, one or more catalytic domains of the Cas7-11protein are mutated to produce a mutated Cas protein which cleaves onlyone DNA strand of a target sequence.

In particular embodiments, the CRISPR-Cas protein may be mutated withrespect to a corresponding wild-type enzyme such that the mutatedCRISPR-Cas protein lacks substantially all DNA cleavage activity. Insome embodiments, a CRISPR-Cas protein may be considered tosubstantially lack all DNA and/or RNA cleavage activity when thecleavage activity of the mutated enzyme is about no more than 25%, 10%,5%, 1%, 0.1%, 0.01%, or less of the nucleic acid cleavage activity ofthe non-mutated form of the enzyme; an example can be when the nucleicacid cleavage activity of the mutated form is nil or negligible ascompared with the non-mutated form.

In certain embodiments of the methods provided herein the CRISPR-Casprotein is a mutated CRISPR-Cas protein which cleaves only one DNAstrand, i.e., a nickase. More particularly, in the context of thepresent disclosure, the nickase ensures cleavage within the non-targetsequence, i.e., the sequence which is on the opposite DNA strand of thetarget sequence and 3′ of the PAM sequence.

In some embodiments, a CRISPR-Cas protein is considered to substantiallylack all DNA cleavage activity when the DNA cleavage activity of themutated enzyme is about no more than 25%, 10%, 5%, 1%, 0.1%, 0.01%, orless of the DNA cleavage activity of the non-mutated form of the enzyme;an example can be when the DNA cleavage activity of the mutated form isnil or negligible as compared with the non-mutated form. In theseembodiments, the CRISPR-Cas protein is used as a generic DNA bindingprotein. The mutations may be artificially introduced mutations or gain-or loss-of-function mutations.

In addition to the mutations described above, the CRISPR-Cas protein maybe additionally modified. As used herein, the term “modified” withregard to a CRISPR-Cas protein generally refers to a CRISPR-Cas proteinhaving one or more modifications or mutations (including pointmutations, truncations, insertions, deletions, chimeras, fusionproteins, etc.) compared to the wild type Cas protein from which it isderived. A modification by truncation can refer to an engineeredtruncation that is based on structure function analysis and notnaturally occurring. By derived is meant that the derived enzyme islargely based, in the sense of having a high degree of sequence homologywith, a wildtype enzyme, but that it has been mutated (modified) in someway as known in the art or as described herein. The modification can befusions of effectors like fluorophore, proteins involved in translationmodulation (e.g., eIF4E, eIF4A, and eIF4G) and proteins involved withepitranscriptomic modulation (e.g., pseudouridine synthase and m6awriter/readers), and splicing factors involved with changing splicing.Cas7-11 could also be used for sensing RNA for diagnostic purposes.

In some embodiments, the C-terminus of the Cas7-11 effector can betruncated. For example, at least 1 amino acid, at least 2 amino acids,at least 3 amino acids, at least 4 amino acids, at least 5 amino acids,at least 6 amino acids, at least 7 amino acids, at least 8 amino acids,at least 9 amino acids, at least 10 amino acids, at least 15 aminoacids, at least 20 amino acids, at least 40 amino acids, at least 50amino acids, at least 60 amino acids, at least 80 amino acids, at least100 amino acids, at least 120 amino acids, at least 140 amino acids, atleast 150 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 250 amino acids, at least 260 amino acids, atleast 300 amino acids, at least 350 amino acids, or any ranges that aremade of any two or more points in the above list may be truncated at theC-terminus of the Cas7-11 effector. For example, up to 120 amino acids,up to 140 amino acids, up to 160 amino acids, up to 180 amino acids, upto 200 amino acids, up to 250 amino acids, up to 300 amino acids, up to350 amino acids, up to 400 amino acids, or any ranges that are made ofany two or more points in the above list may be truncated at theC-terminus of the Cas7-11 effector.

In some embodiments, the N-terminus of the Cas7-11 effector protein maybe truncated. For example, at least 1 amino acid, at least 2 aminoacids, at least 3 amino acids, at least 4 amino acids, at least 5 aminoacids, at least 6 amino acids, at least 7 amino acids, at least 8 aminoacids, at least 9 amino acids, at least 10 amino acids, at least 15amino acids, at least 20 amino acids, at least 40 amino acids, at least50 amino acids, at least 60 amino acids, at least 80 amino acids, atleast 100 amino acids, at least 120 amino acids, at least 140 aminoacids, at least 150 amino acids, at least 160 amino acids, at least 180amino acids, at least 200 amino acids, at least 220 amino acids, atleast 240 amino acids, at least 250 amino acids, at least 260 aminoacids, at least 300 amino acids, at least 350 amino acids, or any rangesthat are made of any two or more points in the above list may betruncated at the N-terminus of the Cas7-11 effector. For examples, up to120 amino acids, up to 140 amino acids, up to 160 amino acids, up to 180amino acids, up to 200 amino acids, up to 250 amino acids, up to 300amino acids, up to 350 amino acids, up to 400 amino acids, or any rangesthat are made of any two or more points in the above list may betruncated at the N-terminus of the Cas7-11 effector.

In some embodiments, both the N- and the C-termini of the Cas7-11effector protein may be truncated. For example, at least 20 amino acidsmay be truncated at the C-terminus of the Cas7-11 effector, and at least20 amino acids, at least 40 amino acids, at least 60 amino acids, atleast 80 amino acids, at least 100 amino acids, at least 120 aminoacids, at least 140 amino acids, at least 160 amino acids, at least 180amino acids, at least 200 amino acids, at least 220 amino acids, atleast 240 amino acids, at least 260 amino acids, at least 300 aminoacids, or at least 350 amino acids may be truncated at the N-terminus ofthe Cas7-11 effector. For example, at least 40 amino acids may betruncated at the C-terminus of the Cas7-11 effector, and at least 20amino acids, at least 40 amino acids, at least 60 amino acids, at least80 amino acids, at least 100 amino acids, at least 120 amino acids, atleast 140 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 260 amino acids, at least 300 amino acids, or atleast 350 amino acids may be truncated at the N-terminus of the Cas7-11effector. For example, at least 60 amino acids may be truncated at theC-terminus of the Cas7-11 effector, and at least 20 amino acids, atleast 40 amino acids, at least 60 amino acids, at least 80 amino acids,at least 100 amino acids, at least 120 amino acids, at least 140 aminoacids, at least 160 amino acids, at least 180 amino acids, at least 200amino acids, at least 220 amino acids, at least 240 amino acids, atleast 260 amino acids, at least 300 amino acids, or at least 350 aminoacids may be truncated at the N-terminus of the Cas7-11 effector. Forexample, at least 80 amino acids may be truncated at the C-terminus ofthe Cas7-11 effector, and at least 20 amino acids, at least 40 aminoacids, at least 60 amino acids, at least 80 amino acids, at least 100amino acids, at least 120 amino acids, at least 140 amino acids, atleast 160 amino acids, at least 180 amino acids, at least 200 aminoacids, at least 220 amino acids, at least 240 amino acids, at least 260amino acids, at least 300 amino acids, or at least 350 amino acids maybe truncated at the N-terminus of the Cas7-11 effector. For example, atleast 100 amino acids may be truncated at the C-terminus of the Cas7-11effector, and at least 20 amino acids, at least 40 amino acids, at least60 amino acids, at least 80 amino acids, at least 100 amino acids, atleast 120 amino acids, at least 140 amino acids, at least 160 aminoacids, at least 180 amino acids, at least 200 amino acids, at least 220amino acids, at least 240 amino acids, at least 260 amino acids, atleast 300 amino acids, or at least 350 amino acids may be truncated atthe N-terminus of the Cas7-11 effector. For example, at least 120 aminoacids may be truncated at the C-terminus of the Cas7-11 effector, and atleast 20 amino acids, at least 40 amino acids, at least 60 amino acids,at least 80 amino acids, at least 100 amino acids, at least 120 aminoacids, at least 140 amino acids, at least 160 amino acids, at least 180amino acids, at least 200 amino acids, at least 220 amino acids, atleast 240 amino acids, at least 260 amino acids, at least 300 aminoacids, or at least 350 amino acids may be truncated at the N-terminus ofthe Cas7-11 effector. For example, at least 140 amino acids may betruncated at the C-terminus of the Cas7-11 effector, and at least 20amino acids, at least 40 amino acids, at least 60 amino acids, at least80 amino acids, at least 100 amino acids, at least 120 amino acids, atleast 140 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 260 amino acids, at least 300 amino acids, or atleast 350 amino acids may be truncated at the N-terminus of the Cas7-11effector. For example, at least 160 amino acids may be truncated at theC-terminus of the Cas7-11 effector, and at least 20 amino acids, atleast 40 amino acids, at least 60 amino acids, at least 80 amino acids,at least 100 amino acids, at least 120 amino acids, at least 140 aminoacids, at least 160 amino acids, at least 180 amino acids, at least 200amino acids, at least 220 amino acids, at least 240 amino acids, atleast 260 amino acids, at least 300 amino acids, or at least 350 aminoacids may be truncated at the N-terminus of the Cas7-11 effector. Forexample, at least 180 amino acids may be truncated at the C-terminus ofthe Cas7-11 effector, and at least 20 amino acids, at least 40 aminoacids, at least 60 amino acids, at least 80 amino acids, at least 100amino acids, at least 120 amino acids, at least 140 amino acids, atleast 160 amino acids, at least 180 amino acids, at least 200 aminoacids, at least 220 amino acids, at least 240 amino acids, at least 260amino acids, at least 300 amino acids, or at least 350 amino acids maybe truncated at the N-terminus of the Cas7-11 effector. For example, atleast 200 amino acids may be truncated at the C-terminus of the Cas7-11effector, and at least 20 amino acids, at least 40 amino acids, at least60 amino acids, at least 80 amino acids, at least 100 amino acids, atleast 120 amino acids, at least 140 amino acids, at least 160 aminoacids, at least 180 amino acids, at least 200 amino acids, at least 220amino acids, at least 240 amino acids, at least 260 amino acids, atleast 300 amino acids, or at least 350 amino acids may be truncated atthe N-terminus of the Cas7-11 effector. For example, at least 220 aminoacids may be truncated at the C-terminus of the Cas7-11 effector, and atleast 20 amino acids, at least 40 amino acids, at least 60 amino acids,at least 80 amino acids, at least 100 amino acids, at least 120 aminoacids, at least 140 amino acids, at least 160 amino acids, at least 180amino acids, at least 200 amino acids, at least 220 amino acids, atleast 240 amino acids, at least 260 amino acids, at least 300 aminoacids, or at least 350 amino acids may be truncated at the N-terminus ofthe Cas7-11 effector. For example, at least 240 amino acids may betruncated at the C-terminus of the Cas7-11 effector, and at least 20amino acids, at least 40 amino acids, at least 60 amino acids, at least80 amino acids, at least 100 amino acids, at least 120 amino acids, atleast 140 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 260 amino acids, at least 300 amino acids, or atleast 350 amino acids may be truncated at the N-terminus of the Cas7-11effector. For example, at least 260 amino acids may be truncated at theC-terminus of the Cas7-11 effector, and at least 20 amino acids, atleast 40 amino acids, at least 60 amino acids, at least 80 amino acids,at least 100 amino acids, at least 120 amino acids, at least 140 aminoacids, at least 160 amino acids, at least 180 amino acids, at least 200amino acids, at least 220 amino acids, at least 240 amino acids, atleast 260 amino acids, at least 300 amino acids, or at least 350 aminoacids may be truncated at the N-terminus of the Cas7-11 effector. Forexample, at least 280 amino acids may be truncated at the C-terminus ofthe Cas7-11 effector, and at least 20 amino acids, at least 40 aminoacids, at least 60 amino acids, at least 80 amino acids, at least 100amino acids, at least 120 amino acids, at least 140 amino acids, atleast 160 amino acids, at least 180 amino acids, at least 200 aminoacids, at least 220 amino acids, at least 240 amino acids, at least 260amino acids, at least 300 amino acids, or at least 350 amino acids maybe truncated at the N-terminus of the Cas7-11 effector. For example, atleast 300 amino acids may be truncated at the C-terminus of the Cas7-11effector, and at least 20 amino acids, at least 40 amino acids, at least60 amino acids, at least 80 amino acids, at least 100 amino acids, atleast 120 amino acids, at least 140 amino acids, at least 160 aminoacids, at least 180 amino acids, at least 200 amino acids, at least 220amino acids, at least 240 amino acids, at least 260 amino acids, atleast 300 amino acids, or at least 350 amino acids may be truncated atthe N-terminus of the Cas7-11 effector. For example, at least 350 aminoacids may be truncated at the C-terminus of the Cas7-11 effector, and atleast 20 amino acids, at least 40 amino acids, at least 60 amino acids,at least 80 amino acids, at least 100 amino acids, at least 120 aminoacids, at least 140 amino acids, at least 160 amino acids, at least 180amino acids, at least 200 amino acids, at least 220 amino acids, atleast 240 amino acids, at least 260 amino acids, at least 300 aminoacids, or at least 350 amino acids may be truncated at the N-terminus ofthe Cas7-11 effector. For example, at least 20 amino acids may betruncated at the N-terminus of the Cas7-11 effector, and at least 20amino acids, at least 40 amino acids, at least 60 amino acids, at least80 amino acids, at least 100 amino acids, at least 120 amino acids, atleast 140 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 260 amino acids, at least 300 amino acids, or atleast 350 amino acids may be truncated at the C-terminus of the Cas7-11effector. For example, at least 40 amino acids may be truncated at theN-terminus of the Cas7-11 effector, and at least 20 amino acids, atleast 40 amino acids, at least 60 amino acids, at least 80 amino acids,at least 100 amino acids, at least 120 amino acids, at least 140 aminoacids, at least 160 amino acids, at least 180 amino acids, at least 200amino acids, at least 220 amino acids, at least 240 amino acids, atleast 260 amino acids, at least 300 amino acids, or at least 350 aminoacids may be truncated at the C-terminus of the Cas7-11 effector. Forexample, at least 60 amino acids may be truncated at the N-terminus ofthe Cas7-11 effector, and at least 20 amino acids, at least 40 aminoacids, at least 60 amino acids, at least 80 amino acids, at least 100amino acids, at least 120 amino acids, at least 140 amino acids, atleast 160 amino acids, at least 180 amino acids, at least 200 aminoacids, at least 220 amino acids, at least 240 amino acids, at least 260amino acids, at least 300 amino acids, or at least 350 amino acids maybe truncated at the C-terminus of the Cas7-11 effector. For example, atleast 80 amino acids may be truncated at the N-terminus of the Cas7-11effector, and at least 20 amino acids, at least 40 amino acids, at least60 amino acids, at least 80 amino acids, at least 100 amino acids, atleast 120 amino acids, at least 140 amino acids, at least 160 aminoacids, at least 180 amino acids, at least 200 amino acids, at least 220amino acids, at least 240 amino acids, at least 260 amino acids, atleast 300 amino acids, or at least 350 amino acids may be truncated atthe C-terminus of the Cas7-11 effector. For example, at least 100 aminoacids may be truncated at the N-terminus of the Cas7-11 effector, and atleast 20 amino acids, at least 40 amino acids, at least 60 amino acids,at least 80 amino acids, at least 100 amino acids, at least 120 aminoacids, at least 140 amino acids, at least 160 amino acids, at least 180amino acids, at least 200 amino acids, at least 220 amino acids, atleast 240 amino acids, at least 260 amino acids, at least 300 aminoacids, or at least 350 amino acids may be truncated at the C-terminus ofthe Cas7-11 effector. For example, at least 120 amino acids may betruncated at the N-terminus of the Cas7-11 effector, and at least 20amino acids, at least 40 amino acids, at least 60 amino acids, at least80 amino acids, at least 100 amino acids, at least 120 amino acids, atleast 140 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 260 amino acids, at least 300 amino acids, or atleast 350 amino acids may be truncated at the C-terminus of the Cas7-11effector. For example, at least 140 amino acids may be truncated at theN-terminus of the Cas7-11 effector, and at least 20 amino acids, atleast 40 amino acids, at least 60 amino acids, at least 80 amino acids,at least 100 amino acids, at least 120 amino acids, at least 140 aminoacids, at least 160 amino acids, at least 180 amino acids, at least 200amino acids, at least 220 amino acids, at least 240 amino acids, atleast 260 amino acids, at least 300 amino acids, or at least 350 aminoacids may be truncated at the C-terminus of the Cas7-11 effector. Forexample, at least 160 amino acids may be truncated at the N-terminus ofthe Cas7-11 effector, and at least 20 amino acids, at least 40 aminoacids, at least 60 amino acids, at least 80 amino acids, at least 100amino acids, at least 120 amino acids, at least 140 amino acids, atleast 160 amino acids, at least 180 amino acids, at least 200 aminoacids, at least 220 amino acids, at least 240 amino acids, at least 260amino acids, at least 300 amino acids, or at least 350 amino acids maybe truncated at the C-terminus of the Cas7-11 effector. For example, atleast 180 amino acids may be truncated at the N-terminus of the Cas7-11effector, and at least 20 amino acids, at least 40 amino acids, at least60 amino acids, at least 80 amino acids, at least 100 amino acids, atleast 120 amino acids, at least 140 amino acids, at least 160 aminoacids, at least 180 amino acids, at least 200 amino acids, at least 220amino acids, at least 240 amino acids, at least 260 amino acids, atleast 300 amino acids, or at least 350 amino acids may be truncated atthe C-terminus of the Cas7-11 effector. For example, at least 200 aminoacids may be truncated at the N-terminus of the Cas7-11 effector, and atleast 20 amino acids, at least 40 amino acids, at least 60 amino acids,at least 80 amino acids, at least 100 amino acids, at least 120 aminoacids, at least 140 amino acids, at least 160 amino acids, at least 180amino acids, at least 200 amino acids, at least 220 amino acids, atleast 240 amino acids, at least 260 amino acids, at least 300 aminoacids, or at least 350 amino acids may be truncated at the C-terminus ofthe Cas7-11 effector. For example, at least 220 amino acids may betruncated at the N-terminus of the Cas7-11 effector, and at least 20amino acids, at least 40 amino acids, at least 60 amino acids, at least80 amino acids, at least 100 amino acids, at least 120 amino acids, atleast 140 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 260 amino acids, at least 300 amino acids, or atleast 350 amino acids may be truncated at the C-terminus of the Cas7-11effector. For example, at least 240 amino acids may be truncated at theN-terminus of the Cas7-11 effector, and at least 20 amino acids, atleast 40 amino acids, at least 60 amino acids, at least 80 amino acids,at least 100 amino acids, at least 120 amino acids, at least 140 aminoacids, at least 160 amino acids, at least 180 amino acids, at least 200amino acids, at least 220 amino acids, at least 240 amino acids, atleast 260 amino acids, at least 300 amino acids, or at least 350 aminoacids may be truncated at the C-terminus of the Cas7-11 effector. Forexample, at least 260 amino acids may be truncated at the N-terminus ofthe Cas7-11 effector, and at least 20 amino acids, at least 40 aminoacids, at least 60 amino acids, at least 80 amino acids, at least 100amino acids, at least 120 amino acids, at least 140 amino acids, atleast 160 amino acids, at least 180 amino acids, at least 200 aminoacids, at least 220 amino acids, at least 240 amino acids, at least 260amino acids, at least 300 amino acids, or at least 350 amino acids maybe truncated at the C-terminus of the Cas7-11 effector. For example, atleast 280 amino acids may be truncated at the N-terminus of the Cas7-11effector, and at least 20 amino acids, at least 40 amino acids, at least60 amino acids, at least 80 amino acids, at least 100 amino acids, atleast 120 amino acids, at least 140 amino acids, at least 160 aminoacids, at least 180 amino acids, at least 200 amino acids, at least 220amino acids, at least 240 amino acids, at least 260 amino acids, atleast 300 amino acids, or at least 350 amino acids may be truncated atthe C-terminus of the Cas7-11 effector. For example, at least 300 aminoacids may be truncated at the N-terminus of the Cas7-11 effector, and atleast 20 amino acids, at least 40 amino acids, at least 60 amino acids,at least 80 amino acids, at least 100 amino acids, at least 120 aminoacids, at least 140 amino acids, at least 160 amino acids, at least 180amino acids, at least 200 amino acids, at least 220 amino acids, atleast 240 amino acids, at least 260 amino acids, at least 300 aminoacids, or at least 350 amino acids may be truncated at the C-terminus ofthe Cas7-11 effector. For example, at least 350 amino acids may betruncated at the N-terminus of the Cas7-11 effector, and at least 20amino acids, at least 40 amino acids, at least 60 amino acids, at least80 amino acids, at least 100 amino acids, at least 120 amino acids, atleast 140 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 260 amino acids, at least 300 amino acids, or atleast 350 amino acids may be truncated at the C-terminus of the Cas7-11effector.

In some embodiments, the Cas7-11 effector comprises a deletion of theINS domain. For example, at least 1 amino acid, at least 2 amino acids,at least 3 amino acids, at least 4 amino acids, at least 5 amino acids,at least 6 amino acids, at least 7 amino acids, at least 8 amino acids,at least 9 amino acids, at least 10 amino acids, at least 15 aminoacids, at least 20 amino acids, at least 40 amino acids, at least 50amino acids, at least 60 amino acids, at least 80 amino acids, at least100 amino acids, at least 120 amino acids, at least 140 amino acids, atleast 150 amino acids, at least 160 amino acids, at least 180 aminoacids, at least 200 amino acids, at least 220 amino acids, at least 240amino acids, at least 250 amino acids, at least 260 amino acids, atleast 300 amino acids, at least 350 amino acids, or any ranges that aremade of any two or more points in the above list of the INS domain maybe deleted.

In some embodiments, the INS domain of the Cas7-11 effector is replacedby a linker. See, e.g., Reddy Chichili, V. P., Kumar, V., & Sivaraman,J., “Linkers in the structural biology of protein-protein interactions,”Protein science: a publication of the Protein Society, 22(2), 153-167(2013); https//doi.org/10.1002/pro.2206, incorporated herewith in itsentirety by reference. For example, the INS domain of the Cas7-11effector may be replaced by a GG, GGG, GS, GGS, GGGS(SEQ ID NO:77),and/or GGGGS(SEQ ID NO:78) linker. For example, the INS domain of theCas7-11 effector may be replaced by a (GG)x(SEQ ID NO:114), (GGG)x(SEQID NO:115), (GGS)x(SEQ ID NO:116), (GGGS)x(SEQ ID NO:117), and/or a(GGGGS)x(SEQ ID NO:118) linker, wherein x is independently 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, or 12. For example, the INS domain of the Cas7-11effector may be replaced by a linker with at least 1 amino acid, atleast 2 amino acids, at least 3 amino acids, at least 4 amino acids, atleast 5 amino acids, at least 6 amino acids, at least 7 amino acids, atleast 8 amino acids, at least 9 amino acids, at least 10 amino acids, atleast 11 amino acids, at least 12 amino acids, at least 13 amino acids,at least 14 amino acids, at least 15 amino acids, at least 16 aminoacids, at least 17 amino acids, at least 18 amino acids, at least 19amino acids, at least 20 amino acids, or any ranges that are made of anytwo or more points in the above list.

The additional modifications of the CRISPR-Cas protein may or may notcause an altered functionality. By means of example, and in particularwith reference to CRISPR-Cas protein, modifications which do not resultin an altered functionality include for instance codon optimization forexpression into a particular host or providing the nuclease with aparticular marker (e.g., for visualization). Modifications with mayresult in altered functionality may also include mutations, includingpoint mutations, insertions, deletions, truncations (including splitnucleases), etc. Fusion proteins may without limitation include forinstance fusions with heterologous domains or functional domains (e.g.,localization signals, catalytic domains, etc.). In certain embodiments,various modifications may be combined (e.g., a mutated nuclease which iscatalytically inactive, and which further is fused to a functionaldomain, such as for instance to induce DNA methylation or anothernucleic acid modification, such as including without limitation a break(e.g., by a different nuclease (domain)), a mutation, a deletion, aninsertion, a replacement, a ligation, a digestion, a break or arecombination). As used herein, “altered functionality” includes withoutlimitation an altered specificity (e.g., altered target recognition,increased (e.g., “enhanced” Cas proteins) or decreased specificity, oraltered PAM recognition), altered activity (e.g., increased or decreasedcatalytic activity, including catalytically inactive nucleases ornickases), and/or altered stability (e.g., fusions with destabilizationdomains). Suitable heterologous domains include without limitation anuclease, a ligase, a repair protein, a methyltransferase, (viral)integrase, a recombinase, a transposase, an argonaute, a cytidinedeaminase, a retron, a group II intron, a phosphatase, a phosphorylase,a sulfurylase, a kinase, a polymerase, an exonuclease, etc. Examples ofall these modifications are known in the art. It will be understood thata “modified” nuclease as referred to herein, and in particular a“modified” Cas or “modified” CRISPR-Cas system or complex preferablystill has the capacity to interact with or bind to the poly-nucleic acid(e.g., in complex with the guide molecule). Such modified Cas proteincan be combined with the deaminase protein or active domain thereof asdescribed herein.

In certain embodiments, CRISPR-Cas protein may comprise one or moremodifications resulting in enhanced activity and/or specificity, such asincluding mutating residues that stabilize the targeted or non-targetedstrand (e.g., eCas9; “Rationally engineered Cas9 nucleases with improvedspecificity”, Slaymaker et al. (2016), Science, 351(6268):84-88,incorporated herewith in its entirety by reference). In certainembodiments, the altered or modified activity of the engineered CRISPRprotein comprises increased targeting efficiency or decreased off-targetbinding. In certain embodiments, the altered activity of the engineeredCRISPR protein comprises modified cleavage activity. In certainembodiments, the altered activity comprises increased cleavage activityas to the target polynucleotide loci. In certain embodiments, thealtered activity comprises decreased cleavage activity as to the targetpolynucleotide loci. In certain embodiments, the altered activitycomprises decreased cleavage activity as to off-target polynucleotideloci. In certain embodiments, the altered or modified activity of themodified nuclease comprises altered helicase kinetics. In certainembodiments, the modified nuclease comprises a modification that altersassociation of the protein with the nucleic acid molecule comprising RNA(in the case of a Cas protein), or a strand of the target polynucleotideloci, or a strand of off-target polynucleotide loci. In an aspect of thedisclosure, the engineered CRISPR protein comprises a modification thatalters formation of the CRISPR complex. In certain embodiments, thealtered activity comprises increased cleavage activity as to off-targetpolynucleotide loci. Accordingly, in certain embodiments, there isincreased specificity for target polynucleotide loci as compared tooff-target polynucleotide loci. In other embodiments, there is reducedspecificity for target polynucleotide loci as compared to off-targetpolynucleotide loci. In certain embodiments, the mutations result indecreased off-target effects (e.g., cleavage or binding properties,activity, or kinetics), such as in case for Cas proteins for instanceresulting in a lower tolerance for mismatches between target and guideRNA. Other mutations may lead to increased off-target effects (e.g.,cleavage or binding properties, activity, or kinetics). Other mutationsmay lead to increased or decreased on-target effects (e.g., cleavage orbinding properties, activity, or kinetics). In certain embodiments, themutations result in altered (e.g., increased or decreased) helicaseactivity, association, or formation of the functional nuclease complex(e.g., CRISPR-Cas complex). In certain embodiments, as described above,the mutations result in an altered PAM recognition, i.e., a differentPAM may be (in addition or in the alternative) be recognized, comparedto the unmodified Cas protein. Particularly preferred mutations includepositively charged residues and/or (evolutionary) conserved residues,such as conserved positively charged residues, in order to enhancespecificity. In certain embodiments, such residues may be mutated touncharged residues, such as alanine. In certain embodiments, suchresidues may be mutated to charged residues, such as arginine andlysine.

Type-III CRISPR-Cas Proteins

The application describes methods using Type-III CRISPR-Cas proteins.This is exemplified herein with Cas7-11, whereby a number of orthologsor homologs have been identified. It will be apparent to the skilledperson that further orthologs or homologs can be identified and that anyof the functionalities described herein may be engineered into otherorthologs, including chimeric enzymes comprising fragments from multipleorthologs.

Computational methods of identifying novel CRISPR-Cas loci are describedin EP3009511 or US2016208243 and may comprise the following steps:detecting all contigs encoding the Cas1 protein; identifying allpredicted protein coding genes within 20 kB of the cas1 gene; comparingthe identified genes with Cas protein-specific profiles and predictingCRISPR arrays; selecting unclassified candidate CRISPR-Cas locicontaining proteins larger than 500 amino acids (>500 aa); analyzingselected candidates using methods such as PSI-BLAST and HHPred to screenfor known protein domains, thereby identifying novel Class 2 CRISPR-Casloci (see also Schmakov et al. 2015, Mol Cell. 60(3):385-97). Inaddition to the above-mentioned steps, additional analysis of thecandidates may be conducted by searching metagenomics databases foradditional homologs. Additionally, or alternatively, to expand thesearch to non-autonomous CRISPR-Cas systems, the same procedure can beperformed with the CRISPR array used as the seed.

In one aspect the detecting all contigs encoding the Cas1 protein isperformed by GenemarkS, a gene prediction program as further describedin “GeneMarkS: a self-training method for prediction of gene starts inmicrobial genomes. Implications for finding sequence motifs inregulatory regions.” John Besemer, Alexandre Lomsadze and MarkBorodovsky, Nucleic Acids Research (2001) 29, pp 2607-2618, hereinincorporated by reference.

In one aspect the identifying all predicted protein coding genes iscarried out by comparing the identified genes with Cas protein-specificprofiles and annotating them according to NCBI Conserved Domain Database(CDD) which is a protein annotation resource that consists of acollection of well-annotated multiple sequence alignment models forancient domains and full-length proteins. These are available asposition-specific score matrices (PSSMs) for fast identification ofconserved domains in protein sequences via RPS-BLAST. CDD contentincludes NCBI-curated domains, which use 3D-structure information toexplicitly define domain boundaries and provide insights intosequence/structure/function relationships, as well as domain modelsimported from a number of external source databases (Pfam, SMART, COG,PRK, TIGRFAM). In a further aspect, CRISPR arrays were predicted using aPILER-CR program which is a public domain software for finding CRISPRrepeats as described in “PILER-CR: fast and accurate identification ofCRISPR repeats,” Edgar, R. C., BMC Bioinformatics, Jan 20; 8:18(2007),herein incorporated by reference.

In a further aspect, the case-by-case analysis is performed usingPSI-BLAST (Position-Specific Iterative Basic Local Alignment SearchTool). PSI-BLAST derives a position-specific scoring matrix (PSSM) orprofile from the multiple sequence alignment of sequences detected abovea given score threshold using protein-protein BLAST. This PSSM is usedto further search the database for new matches and updated forsubsequent iterations with these newly detected sequences. Thus,PSI-BLAST provides a means of detecting distant relationships betweenproteins.

In another aspect, the case-by-case analysis is performed using Hhpred,a method for sequence database searching and structure prediction thatis as easy to use as BLAST or PSI-BLAST and that is at the same timemuch more sensitive in finding remote homologs. In fact, Hhpred'ssensitivity is competitive with the most powerful servers for structureprediction currently available. Hhpred is the first server that is basedon the pairwise comparison of profile hidden Markov models (HMMs).Whereas most conventional sequence search methods search sequencedatabases such as UniProt or the NR, Hhpred searches alignmentdatabases, like Pfam or SMART. This greatly simplifies the list of hitsto a number of sequence families instead of a clutter of singlesequences. All major publicly available profile and alignment databasesare available through Hhpred. Hhpred accepts a single query sequence ora multiple alignment as input. Within only a few minutes it returns thesearch results in an easy-to-read format similar to that of PSI-BLAST.Search options include local or global alignment and scoring secondarystructure similarity. Hhpred can produce pairwise query-templatesequence alignments, merged query-template multiple alignments (e.g.,for transitive searches), as well as 3D structural models calculated bythe MODELLER software from Hhpred alignments.

Deactivated/Inactivated Cas7-11 Proteins

Where the Cas7-11 protein has nuclease activity, the Cas7-11 protein maybe modified to have diminished nuclease activity e.g., nucleaseinactivation of at least 70%, at least 80%, at least 90%, at least 95%,at least 97%, or 100% as compared with the wild type enzyme; or to putin another way, a Cas7-11 enzyme having advantageously about 0% of thenuclease activity of the non-mutated or wild type Cas7-11 enzyme orCRISPR-Cas protein, or no more than about 3% or about 5% or about 10% ofthe nuclease activity of the non-mutated or wild type Cas7-11 enzyme.

Modified Cas7-11 enzymes

In particular embodiments, it is of interest to make use of anengineered Cas7-11 protein as defined herein, such as Cas7-11, whereinthe protein complexes with a nucleic acid molecule comprising RNA toform a CRISPR complex, wherein when in the CRISPR complex, the nucleicacid molecule targets one or more target polynucleotide loci, theprotein comprises at least one modification compared to unmodifiedCas7-11 protein, and wherein the CRISPR complex comprising the modifiedprotein has altered activity as compared to the complex comprising theunmodified Cas7-11 protein. It is to be understood that when referringherein to CRISPR “protein,” the Cas7-11 protein is an unmodified ormodified CRISPR-Cas protein (e.g., having increased or decreased or thesame (or no) enzymatic activity, such as without limitation includingCas7-11. The term “CRISPR protein” may be used interchangeably with“CRISPR-Cas protein”, irrespective of whether the CRISPR protein hasaltered, such as increased or decreased (or no) enzymatic activity,compared to the wild type CRISPR protein.

Computational analysis of the primary structure of Cas7-11 nucleasesreveals 5 distinct domain regions.

Based on the above information, mutants can be generated which lead toinactivation of the enzyme or which modify the double strand nuclease tonickase activity. In alternative embodiments, this information is usedto develop enzymes with reduced off-target effects.

In certain of the above-described Cas7-11 enzymes, the enzyme ismodified by mutation of one or more residues (in the Cas7-like domainsas well as the small subunit).

Orthologs of Cas7-11

The terms “orthologue” (also referred to as “ortholog” herein) and“homologue” (also referred to as “homolog” herein) are well known in theart. By means of further guidance, a “homologue” of a protein as usedherein is a protein of the same species which performs the same or asimilar function as the protein it is a homologue of. Homologousproteins may but need not be structurally related or are only partiallystructurally related. An “orthologue” of a protein as used herein is aprotein of a different species which performs the same or a similarfunction as the protein it is an orthologue of. Orthologous proteins maybut need not be structurally related or are only partially structurallyrelated. Homologs and orthologs may be identified by homology modelling(see, e.g., Greer, Science vol. 228 (1985) 1055, and Blundell et al. EurJ Biochem vol 172(1988), 513) or “structural BLAST” (Dey F, Cliff ZhangQ, Petrey D, Honig B. Toward a “structural BLAST”: using structuralrelationships to infer function. Protein Sci. 2013 April; 22(4):359-66.Doi: 10.1002/pro.2225.). See also Shmakov et al. (2015) for applicationin the field of CRISPR-Cas loci.

The present disclosure encompasses the use of a Cas7-11 effectorprotein, derived from a Cas7-11 locus denoted as subtype III-E. Hereinsuch effector proteins are also referred to as “Cas7-11p”, e.g., aCas7-11 protein (and such effector protein or Cas7-11 protein or proteinderived from a Cas7-11 locus is also called “CRISPR-Cas protein”).

In particular embodiments, the effector protein is a Cas7-11 effectorprotein from an organism from a genus comprising Candidatus Jetteniacaeni, Candidatus Scalindua brodae, Desulfobacteraceae, CandidatusMagnetomorum, Desulfonema Ishimotonii, Candidatus Brocadia,Deltaproteobacteria, Syntrophorhabdaceae, or Nitrospirae.

Delivery Cas7-11 Effector

In some embodiments, the Cas7-11 effector and/or peptide sequence areintroduced into a cell as a nucleic acid encoding each protein. Thenucleic acid introduced into the eukaryotic cell is a plasmid DNA orviral vector. In some embodiments, the Cas7-11 effector and/or peptidesequence are introduced into a cell via a ribonucleoprotein (RNP).

Preferably, delivery is in the form of a vector which may be a viralvector, such as a lenti- or baculo- or adeno-viral/adeno-associatedviral vectors, but other means of delivery are known (such as yeastsystems, microvesicles, gene guns/means of attaching vectors to goldnanoparticles) and are provided. The viral vector may be selected from avariety of families/genera of viruses, including, but not limited toMyoviridae, Siphoviridae, Podoviridae, Corticoviridae, Lipothrixviridae,Poxviridae, Iridoviridae, Adenoviridae, Polyomaviridae,Papillomaviridae, Mimiviridae, Pandoravirusa, Salterprovirusa,Inoviridae, Microviridae, Parvoviridae, Circoviridae, Hepadnaviridae,Caulimoviridae, Retroviridae, Cystoviridae, Reoviridae, Bimaviridae,Totiviridae, Partitiviridae, Filoviridae, Orthomyxoviridae, Deltavirusa,Leviviridae, Picomaviridae, Mamaviridae, Secoviridae, Potyviridae,Caliciviridae, Hepeviridae, Astroviridae, Nodaviridae, Tetraviridae,Luteoviridae, Tombusviridae, Coronaviridae, Arteriviridae, Flaviviridae,Togaviridae, Virgaviridae, Bromoviridae, Tymoviridae, Alphaflexiviridae,Sobemovirusa, Idaeovirusa, and Herpesviridae.

A vector may mean not only a viral or yeast system (for instance, wherethe nucleic acids of interest may be operably linked to and under thecontrol of (in terms of expression, such as to ultimately provide aprocessed RNA) a promoter), but also direct delivery of nucleic acidsinto a host cell. For example, baculoviruses may be used for expressionin insect cells. These insect cells may, in turn be useful for producinglarge quantities of further vectors, such as AAV or lentivirus adaptedfor delivery of the present disclosure. Also envisaged is a method ofdelivering the Cas7-11 effector and/or peptide sequence comprisingdelivering to a cell mRNAs encoding each.

In some embodiments, expression of a nucleic acid sequence encoding theCas7-11 effector and/or peptide sequence may be driven by a promoter. Insome embodiments, a single promoter drives expression of a nucleic acidsequence encoding the Cas7-11 effector. In some embodiments, the Cas7-11effector and guide sequence(s) are operably linked to and expressed fromthe same promoter. In some embodiments, the Cas7-11 and guidesequence(s) are expressed from different promoters. For example, thepromoter(s) can be, but are not limited to, a UBC promoter, a PGKpromoter, an EF1A promoter, a CMV promoter, an EFS promoter, a SV40promoter, and a TRE promoter. The promoter may be a weak or a strongpromoter. The promoter may be a constitutive promoter or an induciblepromoter. In some embodiments, the promoter can also be an AAV ITR, andcan be advantageous for eliminating the need for an additional promoterelement, which can take up space in the vector. The additional spacefreed up by use of an AAV ITR can be used to drive the expression ofadditional elements, such as guide sequences. In some embodiments, thepromoter may be a tissue specific promoter.

In some embodiments, an enzyme coding sequence encoding Cas7-11 effectorand/or peptide sequence is codon-optimized for expression in particularcells, such as eukaryotic cells. The eukaryotic cells may be those of orderived from a particular organism, such as a mammal, including but notlimited to human, mouse, rat, rabbit, dog, or non-human primate. Ingeneral, codon optimization refers to a process of modifying a nucleicacid sequence for enhanced expression in the host cells of interest byreplacing at least one codon (e.g., about or more than about 1, 2, 3, 4,5, 10, 15, 20, 25, 50, or more codons) of the native sequence withcodons that are more frequently or most frequently used in the genes ofthat host cell while maintaining the native amino acid sequence. Variousspecies exhibit particular bias for certain codons of a particular aminoacid. Codon bias (differences in codon usage between organisms) oftencorrelates with the efficiency of translation of messenger RNA (mRNA),which is in turn believed to be dependent on, among other things, theproperties of the codons being translated and the availability ofparticular transfer RNA (tRNA) molecules. The predominance of selectedtRNAs in a cell is generally a reflection of the codons used mostfrequently in peptide synthesis. Accordingly, genes can be tailored foroptimal gene expression in a given organism based on codon optimization.Codon usage tables are readily available, for example, at the “CodonUsage Database”, and these tables can be adapted in a number of ways.See Nakamura, Y., et al. “codon usage tabulated from the internationalDNA sequence databases: status for the year 2000” Nucl. Acids Res.28:292 (2000). Computer algorithms for codon optimizing a particularsequence for expression in a particular host cell are also available,such as Gene Forge (Aptagen; Jacobus, Pa.), are also available. In someembodiments, one or more codons (e.g., 1, 2, 3, 4, 5, 10, 15, 20, 25,50, or more, or all codons) in a sequence encoding a Cas7-11 effectorcorrespond to the most frequently used codon for a particular aminoacid.

In some embodiments, a vector encodes a Cas7-11 effector and/or peptidesequence comprising one or more nuclear localization sequences (NLSs),such as about or more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or moreNLSs. In some embodiments, the Cas7-11 protein comprises about or morethan 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more NLSs at or near theamino-terminus, about or more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,or more NLSs at or near the carboxy-terminus, or a combination of these(e.g., one or more NLS at the amino-terminus and one or more NLS at thecarboxy terminus). When more than one NLS is present, each may beselected independently of the others, such that a single NLS may bepresent in more than one copy and/or in combination with one or moreother NLSs present in one or more copies. In some embodiments, an NLS isconsidered near the N- or C-terminus when the nearest amino acid of theNLS is within about 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 40, 50, or moreamino acids along the polypeptide chain from the N- or C-terminus.Typically, an NLS consists of one or more short sequences of positivelycharged lysines or arginines exposed on the protein surface, bur othertypes of NLS are known. In some embodiments, the NLS is between twodomains, for example between the Cas7-11 effector protein and the viralprotein. The NLS may also be between two functional domains separated orflanked by a glycine-serine linker.

In general, the one or more NLSs are of sufficient strength to driveaccumulation of the Cas7-11 effector and/or peptide sequence in adetectable amount in the nucleus of a eukaryotic cell. In general,strength of nuclear localization activity may derive from the number ofNLSs in the Cas7-11 effector and/or other peptide sequences, theparticular NLS used, or a combination of these factors. Detection ofaccumulation in the nucleus may be performed by any suitable technique.For example, a detectable marker may be fused to the Cas7-11 effectorand/or peptide sequence, such that location within a cell may bevisualized, such as in combination with a means for detecting thelocation of the nucleus (e.g., a stain specific for the nucleus such asDAPI). Examples of detectable markers include fluorescent proteins (suchas green fluorescent proteins, or GFP; RFP; CFP), and epitope tags (HAtag, FLAG tag, SNAP tag). Cell nuclei may also be isolated from cells,the contents of which may then be analyzed by any suitable process fordetecting protein, such as immunohistochemistry, Western blot, or enzymeactivity assay. Accumulation in the nucleus may also be determinedindirectly.

In some aspects, the disclosure provides methods comprising deliveringone or more polynucleotides, such as one or more vectors as describedherein, one or more transcripts thereof, and/or one or proteinstranscribed therefrom, to a host cell. In some aspects, the disclosurefurther provides cells produced by such methods, and organisms (such asanimals, plants, or fungi) comprising or produced from such cells. Insome embodiments, a Cas protein in combination with (and optionallycomplexed) with a guide sequence is delivered to a cell. Conventionalviral and non-viral based gene transfer methods can be used to introducenucleic acids in mammalian cells or target tissues. Such methods can beused to administer nucleic acids encoding a Cas7-11 effector and/or apolypeptide to cells in culture, or in a host organism. Non-viral vectordelivery systems include DNA plasmids, RNA (e.g., a transcript of avector described herein), naked nucleic acid, nucleic acid complexedwith a delivery vehicle, such as a liposome, and ribonucleoprotein.Viral vector delivery systems include DNA and RNA viruses, which haveeither episomal or integrated genomes after delivery to the cell. For areview of gene therapy procedures, see Anderson, Science 256:808-8313(1992); Navel and Felgner, TIBTECH 11:211-217 (1993); Mitani and Caskey,TIBTECH 11:162-166 (1993); Dillon, TIBTECH 11:167-175 (1993); Miller,Nature 357:455-460 (1992); Van Brunt, Biotechnology 6(10):1149-1154(1988); Vigne, Restorative Neurology and Neuroscience 8:35-36 (1995);Kremer and Perricaudet, British Medical Bulletin 51(1):31-44 (1995);Haddada et al., in Current Topics in Microbiology and Immunology,Doerfler and Bohm (eds) (1995); and Yu et al., Gene Therapy 1:13-26(1994), which are incorporated herein by reference in their entirety.

The Cas7-11 effector and/or peptide sequence can be delivered usingadeno-associated virus (AAV), lentivirus, adenovirus, or other viralvector types, or combinations thereof. In some embodiments, one or moreCas7-11 effectors and/or one or more guide RNAs can be packaged into oneor more viral vectors. In some embodiments, the Cas7-11 effector and/orpeptide sequence can be delivered via AAV as a trans-splicing system,similar to Lai et al. (Nature Biotechnology, 2005, DOI:10.1038/nbt1153). In some embodiments, the viral vector is delivered tothe tissue of interest by, for example, an intramuscular injection,while other times the viral delivery is via intravenous, transdermal,intranasal, oral, mucosal, intrathecal, intracranial, or other deliverymethods. Such delivery may be either via a single dose, or multipledoses. One skilled in the art understands that the actual dosage to bedelivered herein may vary greatly depending upon a variety of factors,such as the vector chosen, the target cell, organism, or tissue, thegeneral condition of the subject to be treated, the degree oftransformation/modification sought, the administration route, theadministration mode, the type of transformation/modification sought,etc.

The use of RNA or DNA viral based systems for the delivery of nucleicacids takes advantage of highly evolved processes for targeting a virusto specific cells in the body and trafficking the viral payload to thenucleus. Viral vectors can be administered directly to patients (invivo), or they can be used to treat cells in vitro, and the modifiedcells may optionally be administered to patients (ex vivo). Conventionalviral based systems could include retroviral, lentivirus, adenoviral,adeno-associated and herpes simplex virus vectors for gene transfer.Integration in the host genome is possible with the retrovirus,lentivirus, and adeno-associated virus gene transfer methods, oftenresulting in long term expression of the inserted transgene.Additionally, high transduction efficiencies have been observed in manydifferent cell types and target tissues.

In certain embodiments, delivery of the Cas7-11 and/or peptide sequenceto a cell is non-viral. In certain embodiments, the non-viral deliverysystem is selected from a ribonucleoprotein, cationic lipid vehicle,electroporation, nucleofection, calcium phosphate transfection,transfection through membrane disruption using mechanical shear forces,mechanical transfection, and nanoparticle delivery.

In some embodiments, a host cell is transiently or non-transientlytransfected with one or more vectors described herein. In someembodiments, a cell is transfected as it naturally occurs in a subject.In some embodiments, a cell that is transfected is taken from a subject.In some embodiments, the cell is derived from cells taken from asubject, such as a cell line. Cell lines are available from a variety ofsources known to those with skill in the art (see, e.g., the AmericanType Culture Collection (ATCC) (Manassas, VA). In some embodiments, acell transfected with one or more vectors described herein is used toestablish a new cell line comprising one or more vector-derivedsequences.

Guide Molecules

The system may comprise a guide molecule. The guide molecule maycomprise a guide sequence. In certain cases, the guide sequence may belinked to a direct repeat sequence. In some cases, the system maycomprise a nucleotide sequence encoding the guide molecule. The guidemolecule may form a complex with the dead Cas7-11 protein and directsthe complex to bind the target RNA sequence at one or more codonsencoding an amino acid that is post-translationally modified. The guidesequence may be capable of hybridizing with a target RNA sequencecomprising an Adenine or Cytidine encoding said amino acid to form anRNA duplex, wherein said guide sequence comprises a non-pairingnucleotide at a position corresponding to said Adenine or Cytidineresulting in a mismatch in the RNA duplex formed. The guide sequence maycomprise one or more mismatch corresponding to different adenosine sitesin the target sequence. In certain cases, guide sequence may comprisemultiple mismatches corresponding to different adenosine sites in thetarget sequence. In cases where two guide molecules are used, the guidesequence of each of the guide molecules may comprise a mismatchcorresponding to a different adenosine site in the target sequence.

In general, a CRISPR system is characterized by elements that promotethe formation of a CRISPR complex at the site of a target sequence. Inthe context of formation of a CRISPR complex, “target sequence” refersto a sequence to which a guide sequence is designed to havecomplementarity, where hybridization between a target DNA sequence and aguide sequence promotes the formation of a CRISPR complex.

In certain embodiments, the target sequence should be associated with aPAM (protospacer adjacent motif) or PFS (protospacer flanking sequenceor site); that is, a short sequence recognized by the CRISPR complex.Depending on the nature of the CRISPR-Cas protein, the target sequenceshould be selected such that its complementary sequence in the DNAduplex (also referred to herein as the non-target sequence) is upstreamor downstream of the PAM. The precise sequence and length requirementsfor the PAM differ depending on the Cas7-11 protein used, but PAMs aretypically 2-8 base pair sequences adjacent the protospacer (that is, thetarget sequence). Examples of the natural PAM sequences for differentCas7-11 orthologues are provided herein below and the skilled personwill be able to identify further PAM sequences for use with a givenCas7-11 protein. In certain embodiments, the Cas7-11 protein has beenmodified to recognize a non-natural PAM, such as recognizing a PAMhaving a sequence or comprising a sequence YCN, YCV, AYV, TYV, RYN, RCN,TGYV(SEQ ID NO:79), NTTN(SEQ ID NO:80), TN, TRTN(SEQ ID NO:81), TYTV(SEQID NO:82), TYCT(SEQ ID NO:83), TYCN(SEQ ID NO:84), TRTN(SEQ ID NO:81),NTTN(SEQ ID NO:80), TACT(SEQ ID NO:85), TYCC(SEQ ID NO:86), TRTC(SEQ IDNO:87), TATV(SEQ ID NO:88), NTTV(SEQ ID NO:89), TTV, TSTG(SEQ ID NO:90),TVTS(SEQ ID NO:91), TYYS(SEQ ID NO:92), TCYS(SEQ ID NO:93), TBYS(SEQ IDNO:94), TCYS(SEQ ID NO:93), TNYS(SEQ ID NO:95), TYYS(SEQ ID NO:92),TNTN(SEQ ID NO:96), TSTG(SEQ ID NO:90), TTCC(SEQ ID NO:97), TCCC(SEQ IDNO:98), TATC(SEQ ID NO:99), TGTG(SEQ ID NO:100), TCTG(SEQ ID NO:101),TYCV(SEQ ID NO:102), or TCTC(SEQ ID NO:103).

The terms “guide molecule” and “guide RNA” are used interchangeablyherein to refer to RNA-based molecules that are capable of forming acomplex with a CRISPR-Cas protein and comprises a guide sequence havingsufficient complementarity with a target nucleic acid sequence tohybridize with the target nucleic acid sequence and directsequence-specific binding of the complex to the target nucleic acidsequence. The guide molecule or guide RNA specifically encompassesRNA-based molecules having one or more chemically modifications (e.g.,by chemical linking two ribonucleotides or by replacement of one or moreribonucleotides with one or more deoxyribonucleotides), as describedherein.

As used herein, the term “guide sequence” in the context of a CRISPR-Cassystem, comprises any polynucleotide sequence having sufficientcomplementarity with a target nucleic acid sequence to hybridize withthe target nucleic acid sequence and direct sequence-specific binding ofa nucleic acid-targeting complex to the target nucleic acid sequence. Inthe context of the present disclosure the target nucleic acid sequenceor target sequence is the sequence comprising the target adenosine to bedeaminated also referred to herein as the “target adenosine”. In someembodiments, except for the intended dA-C mismatch, the degree ofcomplementarity, when optimally aligned using a suitable alignmentalgorithm, is about or more than about 50%, 60%, 75%, 80%, 85%, 90%,95%, 97.5%, 99%, or more. Optimal alignment may be determined with theuse of any suitable algorithm for aligning sequences, non-limitingexample of which include the Smith-Waterman algorithm, theNeedleman-Wunsch algorithm, algorithms based on the Burrows-WheelerTransform (e.g., the Burrows Wheeler Aligner), ClustalW, ClustalX, BLAT,Novoalign (Novocraft Technologies; available at www.novocraft.com),ELAND (Illumina, San Diego, CA), SOAP (available atsoap.genomics.org.cn), and Maq (available at maq.sourceforge.net). Theability of a guide sequence (within a nucleic acid-targeting guide RNA)to direct sequence-specific binding of a nucleic acid-targeting complexto a target nucleic acid sequence may be assessed by any suitable assay.For example, the components of a nucleic acid-targeting CRISPR systemsufficient to form a nucleic acid-targeting complex, including the guidesequence to be tested, may be provided to a host cell having thecorresponding target nucleic acid sequence, such as by transfection withvectors encoding the components of the nucleic acid-targeting complex,followed by an assessment of preferential targeting (e.g., cleavage)within the target nucleic acid sequence, such as by Surveyor assay asdescribed herein. Similarly, cleavage of a target nucleic acid sequence(or a sequence in the vicinity thereof) may be evaluated in a test tubeby providing the target nucleic acid sequence, components of a nucleicacid-targeting complex, including the guide sequence to be tested and acontrol guide sequence different from the test guide sequence, andcomparing binding or rate of cleavage at or in the vicinity of thetarget sequence between the test and control guide sequence reactions.Other assays are possible, and will occur to those skilled in the art. Aguide sequence, and hence a nucleic acid-targeting guide RNA may beselected to target any target nucleic acid sequence.

In some embodiments, the guide molecule comprises a guide sequence thatis designed to have at least one mismatch with the target sequence, suchthat an RNA duplex formed between the guide sequence and the targetsequence comprises a non-pairing C in the guide sequence opposite to thetarget A for deamination on the target sequence. In some embodiments,aside from this A-C mismatch, the degree of complementarity, whenoptimally aligned using a suitable alignment algorithm, is about or morethan about 50%, 60%, 75%, 80%, 85%, 90%, 95%, 97.5%, 99%, or more. Insome cases, the distance between the non-pairing C and the 5′ end of theguide sequence is from about 10 to about 50, e.g., from about 10 toabout 20, from about 15 to about 25, from about 20 to about 30, fromabout 25 to about 35, from about 30 to about 40, from about 35 to about45, or from about 40 to about 50 nucleotides (nt) in length. In certainexample. In some cases, the distance between the non-pairing C and the3′ end of the guide sequence is from about 10 to about 50, e.g., fromabout 10 to about 20, from about 15 to about 25, from about 20 to about30, from about 25 to about 35, from about 30 to about 40, from about 35to about 45, or from about 40 to about 50 nucleotides (nt) in length. Inone example, the distance between the non-pairing C and the 5′ end ofsaid guide sequence is from about 20 to about 30 nucleotides.

In certain embodiments, the guide sequence or spacer length of the guidemolecules is from 15 to 50 nt. In certain embodiments, the spacer lengthof the guide RNA is at least 15 nucleotides. In certain embodiments, thespacer length is from 15 to 17 nt, e.g., 15, 16, or 17 nt, from 17 to 20nt, e.g., 17, 18, 19, or 20 nt, from 20 to 24 nt, e.g., 20, 21, 22, 23,or 24 nt, from 23 to 25 nt, e.g., 23, 24, or 25 nt, from 24 to 27 nt,e.g., 24, 25, 26, or 27 nt, from 27-30 nt, e.g., 27, 28, 29, or 30 nt,from 30-35 nt, e.g., 30, 31, 32, 33, 34, or 35 nt, or 35 nt or longer.In certain example embodiment, the guide sequence is 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,92, 93, 94, 95, 96, 97, 98, 99, or 100 nt.

In some embodiments, the guide sequence has a length from about 10 toabout 100, e.g., from about 20 to about 60, from about 20 to about 55,from about 20 to about 53, from about 25 to about 53, from about 29 toabout 53, from about 20 to about 30, from about 25 to about 35, fromabout 30 to about 40, from about 35 to about 45, from about 40 to about50, from about 45 to about 55, from about 50 to about 60, from about 55to about 65, from about 60 to about 70, from about 70 to about 80, fromabout 80 to about 90, or from about 90 to about 100 nucleotides (nt)long that is capable of forming an RNA duplex with a target sequence. Incertain example, the guide sequence has a length from about 20 to about53 nt capable of forming said RNA duplex with said target sequence. Incertain example, the guide sequence has a length from about 25 to about53 nt capable of forming said RNA duplex with said target sequence. Incertain example, the guide sequence has a length from about 29 to about53 nt capable of forming said RNA duplex with said target sequence. Incertain example, the guide sequence has a length from about 40 to about50 nt capable of forming said RNA duplex with said target sequence. Insome examples, the guide sequence comprises a non-pairing Cytosine at aposition corresponding to said Adenine resulting in an A-C mismatch inthe RNA duplex formed. The guide sequence is selected so as to ensurethat it hybridizes to the target sequence comprising the adenosine to bedeaminated.

In some embodiments, the guide sequence is about 10 nt to about 100 ntlong and hybridizes to the target DNA strand to form an almost perfectlymatched duplex, except for having a dA-C mismatch at the targetadenosine site. Particularly, in some embodiments, the dA-C mismatch islocated close to the center of the target sequence (and thus the centerof the duplex upon hybridization of the guide sequence to the targetsequence), thereby restricting the nucleotide deaminase to a narrowediting window (e.g., about 4 bp wide). In some embodiments, the targetsequence may comprise more than one target adenosine to be deaminated.In further embodiments, the target sequence may further comprise one ormore dA-C mismatch 3′ to the target adenosine site. In some embodiments,to avoid off-target editing at an unintended Adenine site in the targetsequence, the guide sequence can be designed to comprise a non-pairingGuanine at a position corresponding to said unintended Adenine tointroduce a dA-G mismatch, which is catalytically unfavorable forcertain nucleotide deaminases such as ADAR1 and ADAR2. See Wong et al.,RNA 7:846-858 (2001), which is incorporated herein by reference in itsentirety.

In some embodiments, the sequence of the guide molecule (direct repeatand/or spacer) is selected to reduce the degree secondary structurewithin the guide molecule. In some embodiments, about or less than about75%, 50%, 40%, 30%, 25%, 20%, 15%, 10%, 5%), 1%), or fewer of thenucleotides of the nucleic acid-targeting guide RNA participate inself-complementary base pairing when optimally folded. Optimal foldingmay be determined by any suitable polynucleotide folding algorithm. Someprograms are based on calculating the minimal Gibbs free energy. Anexample of one such algorithm is mFold, as described by Zuker andStiegler (Nucleic Acids Res. 9 (1981), 133-148). Another example foldingalgorithm is the online webserver RNAfold, developed at Institute forTheoretical Chemistry at the University of Vienna, using the centroidstructure prediction algorithm (see e.g., A. R. Gruber et al., 2008,Cell 106(1): 23-24; and P A Carr and G M Church, 2009, NatureBiotechnology 27(12): 1151-62).

In some embodiments, it is of interest to reduce the susceptibility ofthe guide molecule to RNA cleavage, such as to cleavage by Cas7-11.Accordingly, in particular embodiments, the guide molecule is adjustedto avoid cleavage by Cas7-11 or other RNA-cleaving enzymes.

In some embodiments, the guide molecule is modified, e.g., by one ormore aptamer(s) designed to improve guide molecule delivery, includingdelivery across the cellular membrane, to intracellular compartments, orinto the nucleus. Such a structure can include, either in addition tothe one or more aptamer(s) or without such one or more aptamer(s),moiety(ies) so as to render the guide molecule deliverable, inducible orresponsive to a selected effector. The disclosure accordinglycomprehends a guide molecule that responds to normal or pathologicalphysiological conditions, including without limitation pH, hypoxia, O₂concentration, temperature, protein concentration, enzymaticconcentration, lipid structure, light exposure, mechanical disruption(e.g., ultrasound waves), magnetic fields, electric fields, orelectromagnetic radiation.

Adenosine Deaminase

The system may further comprise an adenosine deaminase or catalyticdomain thereof. The adenosine deaminase protein or catalytic domainthereof deaminates an Adenine or Cytidine at the one or more codonsthereby changing the codon to encode for an amino acid that is notpost-translationally modified. The term “adenosine deaminase” or“adenosine deaminase protein” as used herein refers to a protein, apolypeptide, or one or more functional domain(s) of a protein or apolypeptide that is capable of catalyzing a hydrolytic deaminationreaction that converts an adenine (or an adenine moiety of a molecule)to a hypoxanthine (or a hypoxanthine moiety of a molecule), as shownbelow. In some embodiments, the adenine-containing molecule is anadenosine (A), and the hypoxanthine-containing molecule is an inosine(I). The adenine-containing molecule can be deoxyribonucleic acid (DNA)or ribonucleic acid (RNA).

According to the present disclosure, adenosine deaminases that can beused in connection with the present disclosure include, but are notlimited to, members of the enzyme family known as adenosine deaminasesthat act on RNA (ADARs), members of the enzyme family known as adenosinedeaminases that act on tRNA (ADATs), and other adenosine deaminasedomain-containing (AD AD) family members. According to the presentdisclosure, the adenosine deaminase is capable of targeting adenine inan RNA/DNA and RNA duplexes. Indeed, Zheng et al. (Nucleic Acids Res.2017, 45(6): 3369-3377) demonstrate that ADARs can carry out adenosineto inosine editing reactions on RNA/DNA and RNA/RNA duplexes. Theadenosine deaminase can be modified to increase its ability to edit DNAin an RNA/DNA RNA duplex.

In some embodiments, the adenosine deaminase is derived from one or moremetazoa species, including but not limited to, mammals, birds, frogs,squids, fish, flies, and worms. In some embodiments, the adenosinedeaminase is a human, cephalopod (e.g., squid) or Drosophila adenosinedeaminase. In certain examples, the adenosine deaminase is a humanadenosine deaminase. In certain examples, the adenosine deaminase is acephalopod adenosine deaminase. In certain examples, the adenosinedeaminase is a Drosophila adenosine deaminase.

Cytidine Deaminase

The term “cytidine deaminase” or “cytidine deaminase protein” as usedherein refers to a protein, a polypeptide, or one or more functionaldomain(s) of a protein or a polypeptide that is capable of catalyzing ahydrolytic deamination reaction that converts a cytosine (or a cytosinemoiety of a molecule) to an uracil (or an uracil moiety of a molecule),as shown below. In some embodiments, the cytosine-containing molecule isa cytidine (C), and the uracil-containing molecule is a uridine (U). Thecytosine-containing molecule can be deoxyribonucleic acid (DNA) orribonucleic acid (RNA).

According to the present disclosure, cytidine deaminases that can beused in connection with the present disclosure include, but are notlimited to, members of the enzyme family known as apolipoprotein BmRNA-editing complex (APOBEC) family deaminase, an activation-induceddeaminase (AID), or a cytidine deaminase 1 (CDA1). In particularembodiments, the deaminase in an APOBEC 1 deaminase, an APOBEC2deaminase, an APOBEC3A deaminase, an APOBEC3B deaminase, an APOBEC3Cdeaminase, and APOBEC3D deaminase, an APOBEC3E deaminase, an APOBEC3Fdeaminase an APOBEC3G deaminase, an APOBEC3H deaminase, or an APOBEC4deaminase. The cytidine deaminase can be modified to increase itsability to edit DNA in an RNA/DNAn RNA duplex.

In some embodiments, the cytidine deaminase is derived from one or moremetazoa species, including but not limited to, mammals, birds, frogs,squids, fish, flies, and worms. In some embodiments, the cytidinedeaminase is a human, primate, cow, dog, rat, or mouse cytidinedeaminase.

CD (cytidine deaminase)-functionalized CRISPR system for RNA editing canbe used for C to U conversions. In some embodiments, the cytidinedeaminase protein or catalytic domain thereof is a human, rat or lampreycytidine deaminase protein or catalytic domain thereof. In someembodiments, the cytidine deaminase protein or catalytic domain thereofis an apolipoprotein B mRNA-editing complex (APOBEC) family deaminase,an activation-induced deaminase (AID), or a cytidine deaminase 1 (CDA1).In some embodiments, the cytidine deaminase protein or catalytic domainthereof is an APOBEC1 deaminase comprising one or more mutationscorresponding to W90A, W90Y, R118A, H121R, H122R, R126A, R126E, or R132Ein rat APOBEC1, or an APOBEC3G deaminase comprising one or moremutations corresponding to W285A, W285Y, R313A, D316R, D317R, R320A,R320E, or R326E in human APOBEC3G. In some embodiments, the cytidinedeaminase protein or catalytic domain thereof is delivered together withan uracil glycosylase inhibitor (UGI), where said UGI is covalentlylinked to said cytidine deaminase protein or catalytic domain thereofand/or said catalytically inactive Cas7-11 protein.

Cas7-11-APOBEC fusions can perform C-to-U editing of RNA. APOBECsubstrates are ssRNA and the Cas7-11-APOBEC can therefore target regionsof the RNA around the guide/target duplex. Cas7-11-APOBEC fusions canperform C to U knockdown via stop codon introduction. In addition tocorrecting pathogenic U to C mutations that arise during the cellularlife cycle, Cas7-11-APOBEC fusions can lead to the introduction of stopcodons by converting a CAA, CGA, or CAG to TAA, TGA, or TAG,respectively. APOBEC orthologs in fusion with Cas7-11 can increase theefficiency of C-to-U editing or can allow for additional types of baseconversions. Mutating the APOBEC from the Cas7-11-APOBEC can lead tofusions with specific dsRNA activity, base flip activity and increasedactivity.

Ortholog Truncations

Table 1 below shows examples of truncations of Cas7-11 orthologs.

TABLE 1 Residues replaced with GGGS Start End AA of WT AA position AAposition AA of Cas7-11S DiCas7-11S 1601 Met 979 Ser 1293 1291 SbCas7-11S1722 Met 1032 Ser 1387 1371 HvsCas7-11S 1528 Met 955 Ser 1240 1247CjcCas7-11S 1812 Leu 1109 Ser 1406 1519 Correspond to amino acid rangespredicted for truncations that can make the orthologs smaller yet stillfunctional. The truncated orthologs can be useful for deliveryapplications, like AAV, that have packaging size limits. “AA of WT”denotes the length of the unmodified WT ortholog. “AA of Cas7-11S”denotes the length of the ortholog after the proposed deletion betweenthe “Start” and “End” positions and insertion of a GGGS linker. The GGGSlinker helps maintain flexibility of the fused protein domains.

Based on conservation and structural homology to the DiCas7-11structure, predictions can be made in the four orthologs shown in FIG.24 and in Table 1 above for non-functional regions to truncate out,where the orthologs retain catalytic RNA cleavage activity. The columnlabelled START signifies the amino acid at the beginning of the proposedtruncation and the column labelled END signifies the amino acid at theend of the truncation. By subtracting those two numbers, the length ofthe truncation can be determined. The column labelled AA of WT for eachortholog refers to the natural length of the ortholog in amino acids.For example, the first entry, DiCas7-11, refers to the non-truncatedform, which is 1601 amino acids. After the indicated truncation fromposition 979 to 1293 and the addition of the GGGS(SEQ ID NO:77) linker,the modified protein will be 1291 amino acids in length, as indicated inthe column AA of Cas7-11S.

The GGGS(SEQ ID NO:77) linker is used to replace these regions becauseit is a small, flexible linker that ensures that the truncation isfunctional (i.e., can bind a crRNA and target RNA for cleavage).

The resulting truncated orthologs are easier to package for delivery tocells because they are 285-355 amino acids shorter in length and arestill predicted to retain RNA knockdown and RNA binding function basedon the DiCas7-11S truncation.

In some embodiments, the truncation could be made without inserting theGGGS(SEQ ID NO:77) linker. In addition, other linkers could be used orbe placed in other domains. Other possible linker sequences, include,but are not limited to:

GS (SEQ ID NO: 104) GSGGGGS (SEQ ID NO: 105) GGGGSGGGGSGGGGS(SEQ ID NO: 106) EAAAK (SEQ ID NO: 107) EAAAKEAAAKEAAAK (SEQ ID NO: 108)GGSGGSGGSGGSGGSGGS (SEQ ID NO: 109) SGSETPGTSESATPES

Residue Mutations

Amino acid residues that are located near or cRNA or the target RNA canbe varied also. Table 2 shows examples of amino acid residue mutationsto boost the activity of DiCas7-11. The column labeled AA shows theidentity of the residue and the position column indicates the positionof the amino acid in DiCas7-11. In some embodiments, one or moreindividual or multiple residues can be mutated to another amino acidresidue or multiple residues. In some embodiments, the individual aminoacid is mutated to an arginine or a lysine.

TABLE 2 Mutations to Arg or Lys might increase the target RNA bindingand cleavage seems more feasible DiCas7-11 AA position DiCas7-11 Glu 280DiCas7-11 Tyr 281 DiCas7-11 His 306 DiCas7-11 Asp 307 DiCas7-11 Asp 311DiCas7-11 Tyr 313 DiCas7-11 Tyr 360 DiCas7-11 Glu 513 DiCas7-11 Asp 980DiCas7-11 Asp 982 DiCas7-11 Asp 989 DiCas7-11 Asp 1164 DiCas7-11 Asn1247 DiCas7-11 Glu 1392 DiCas7-11 Asn 1441 DiCas7-11 Asp 1568 DiCas7-11Asp 1581 Listed are residues that can modulate activity and/or RNAbinding. The residues are highlighted in the structure model shown inFIG. 24.

EXAMPLES

While several experimental Examples are contemplated, these Examples areintended to be non-limiting.

Materials

Table 3 below shows examples of protein amino acid sequences (N-Cterminal).

TABLE 3 SEQ ID NO/ Description Sequence SEQ IDMTTTMKISIEFLEPFRMTKWQESTRRNKNNKEFVRGQAFARWHRNKK NO: 1DNTKGRPYITGTLLRSAVIRSAENLLTLSDGKISEKTCCPGKFDTEDKD huDiCas7-RLLQLRQRSTLRWTDKNPCPDNAETYCPFCELLGRSGNDGKKAEKKD 11WRFRIHFGNLSLPGKPDFDGPKAIGSQRVLNRVDFKSGKAHDFFKAYEVDHTRFPRFEGEITIDNKVSAEARKLLCDSLKFTDRLCGALCVIRFDEYTPAADSGKQTENVQAEPNANLAEKTAEQIISILDDNKKTEYTRLLADAIRSLRRSSKLVAGLPKDHDGKDDHYLWDIGKKKKDENSVTIRQILTTSADTKELKNAGKWREFCEKLGEALYLKSKDMSGGLKITRRILGDAEFHGKPDRLEKSRSVSIGSVLKETVVCGELVAKTPFFFGAIDEDAKQTDLQVLLTPDNKYRLPRSAVRGILRRDLQTYFDSPCNAELGGRPCMCKTCRIMRGITVMDARSEYNAPPEIRHRTRINPFTGTVAEGALFNMEVAPEGIVFPFQLRYRGSEDGLPDALKTVLKWWAEGQAFMSGAASTGKGRFRMENAKYETLDLSDENQRNDYLKNWGWRDEKGLEELKKRLNSGLPEPGNYRDPKWHEINVSIEMASPFINGDPIRAAVDKRGTDVVTFVKYKAEGEEAKPVCAYKAESFRGVIRSAVARIHMEDGVPLTELTHSDCECLLCQIFGSEYEAGKIRFEDLVFESDPEPVTFDHVAIDRFTGGAADKKKFDDSPLPGSPARPLMLKGSFWIRRDVLEDEEYCKALGKALADVNNGLYPLGGKSAIGYGQVKSLGIKGDDKRISRLMNPAFDETDVAVPEKPKTDAEVRIEAEKVYYPHYFVEPHKKVEREEKPCGHQKFHEGRLTGKIRCKLITKTPLIVPDTSNDDFFRPADKEARKEKDEYHKSYAFFRLHKQIMIPGSELRGMVSSVYETVTNSCFRIFDETKRLSWRMDADHQNVLQDFLPGRVTADGKHIQKFSETARVPFYDKTQKHFDILDEQEIAGEKPVRMWVKRFIKRLSLVDPAKHPQKKQDNKWKRRKEGIATFIEQKNGSYYFNVVTNNGCTSFHLWHKPDNFDQEKLEGIQNGEKLDCWVRDSRYQKAFQEIPENDPDGWECKEGYLHVVGPSKVEFSDKKGDVINNFQGTLPSVPNDWKTIRTNDFKNRKRKNEPVFCCEDDKGNYYTMAKYCETFFFDLKENEEYEIPEKARIKYKELLRVYNNNPQAVPESVFQSRVARENVEKLKSGDLVYFKHNEKYVEDIVPVRISRTVDDRMIGKRMSADLRPCHGDWVEDGDLSALNAYPEKRLLLRHPKGLCPACRLFGTGSYKGRVRFGFASLENDPEWLIPGKNPGDPFHGGPVMLSLLERPRPTWSIPGSDNKFKVPGRKFYVHHHAWKTIKDGNHPTTGKAIEQSPNNRTVEALAGGNSFSFEIAFENLKEWELGLLIHSLQLEKGLAHKLGMAKSMGFGSVEIDVESVRLRKDWKQWRNGNSEIPNWLGKGFAKLKEWFRDELDFIENLKKLLWFPEGDQAPRVCYPMLRKKDDPNGNSGYEELKDGEFKKEDRQKKLTTPWTPWA SEQ IDMTTTMKISIEFLEPFRMTKWQESTRRNKNNKEFVRGQAFARWHRNKK NO: 2DNTKGRPYITGTLLRSAVIRSAENLLTLSDGKISEKTCCPGKFDTEDKD huDiCas7-RLLQLRQRSTLRWTDKNPCPDNAETYCPFCELLGRSGNDGKKAEKKD 11-WRFRIHFGNLSLPGKPDFDGPKAIGSQRVLNRVDFKSGKAHDFFKAYE R978-VDHTRFPRFEGEITIDNKVSAEARKLLCDSLKFTDRLCGALCVIRFDEY GGGS-TPAADSGKQTENVQAEPNANLAEKTAEQIISILDDNKKTEYTRLLADAI R1294RSLRRSSKLVAGLPKDHDGKDDHYLWDIGKKKKDENSVTIRQILTTSADTKELKNAGKWREFCEKLGEALYLKSKDMSGGLKITRRILGDAEFHGKPDRLEKSRSVSIGSVLKETVVCGELVAKTPFFFGAIDEDAKQTDLQVLLTPDNKYRLPRSAVRGILRRDLQTYFDSPCNAELGGRPCMCKTCRIMRGITVMDARSEYNAPPEIRHRTRINPFTGTVAEGALFNMEVAPEGIVFPFQLRYRGSEDGLPDALKTVLKWWAEGQAFMSGAASTGKGRFRMENAKYETLDLSDENQRNDYLKNWGWRDEKGLEELKKRLNSGLPEPGNYRDPKWHEINVSIEMASPFINGDPIRAAVDKRGTDVVTFVKYKAEGEEAKPVCAYKAESFRGVIRSAVARIHMEDGVPLTELTHSDCECLLCQIFGSEYEAGKIRFEDLVFESDPEPVTFDHVAIDRFTGGAADKKKFDDSPLPGSPARPLMLKGSFWIRRDVLEDEEYCKALGKALADVNNGLYPLGGKSAIGYGQVKSLGIKGDDKRISRLMNPAFDETDVAVPEKPKTDAEVRIEAEKVYYPHYFVEPHKKVEREEKPCGHQKFHEGRLTGKIRCKLITKTPLIVPDTSNDDFFRPADKEARKEKDEYHKSYAFFRLHKQIMIPGSELRGMVSSVYETVTNSCFRIFDETKRLSWRGGGSRTVDDRMIGKRMSADLRPCHGDWVEDGDLSALNAYPEKRLLLRHPKGLCPACRLFGTGSYKGRVRFGFASLENDPEWLIPGKNPGDPFHGGPVMLSLLERPRPTWSIPGSDNKFKVPGRKFYVHHHAWKTIKDGNHPTTGKAIEQSPNNRTVEALAGGNSFSFEIAFENLKEWELGLLIHSLQLEKGLAHKLGMAKSMGFGSVEIDVESVRLRKDWKQWRNGNSEIPNWLGKGFAKLKEWFRDELDFIENLKKLLWFPEGDQAPRVCYPMLRKKDDPNGNSGYEELKDGEFKKEDRQKKLTTPWTPW A SEQ IDMTTTMKISIEFLEPFRMTKWQESTRRNKNNKEFVRGQAFARWHRNKK NO: 3DNTKGRPYITGTLLRSAVIRSAENLLTLSDGKISEKTCCPGKFDTEDKD huDiCas7-RLLQLRQRSTLRWTDKNPCPDNAETYCPFCELLGRSGNDGKKAEKKD 11-WRFRIHFGNLSLPGKPDFDGPKAIGSQRVLNRVDFKSGKAHDFFKAYE S1006-VDHTRFPRFEGEITIDNKVSAEARKLLCDSLKFTDRLCGALCVIRFDEY GGGS-TPAADSGKQTENVQAEPNANLAEKTAEQIISILDDNKKTEYTRLLADAI D1221RSLRRSSKLVAGLPKDHDGKDDHYLWDIGKKKKDENSVTIRQILTTSADTKELKNAGKWREFCEKLGEALYLKSKDMSGGLKITRRILGDAEFHGKPDRLEKSRSVSIGSVLKETVVCGELVAKTPFFFGAIDEDAKQTDLQVLLTPDNKYRLPRSAVRGILRRDLQTYFDSPCNAELGGRPCMCKTCRIMRGITVMDARSEYNAPPEIRHRTRINPFTGTVAEGALFNMEVAPEGIVFPFQLRYRGSEDGLPDALKTVLKWWAEGQAFMSGAASTGKGRFRMENAKYETLDLSDENQRNDYLKNWGWRDEKGLEELKKRLNSGLPEPGNYRDPKWHEINVSIEMASPFINGDPIRAAVDKRGTDVVTFVKYKAEGEEAKPVCAYKAESFRGVIRSAVARIHMEDGVPLTELTHSDCECLLCQIFGSEYEAGKIRFEDLVFESDPEPVTFDHVAIDRFTGGAADKKKFDDSPLPGSPARPLMLKGSFWIRRDVLEDEEYCKALGKALADVNNGLYPLGGKSAIGYGQVKSLGIKGDDKRISRLMNPAFDETDVAVPEKPKTDAEVRIEAEKVYYPHYFVEPHKKVEREEKPCGHQKFHEGRLTGKIRCKLITKTPLIVPDTSNDDFFRPADKEARKEKDEYHKSYAFFRLHKQIMIPGSELRGMVSSVYETVTNSCFRIFDETKRLSWRMDADHQNVLQDFLPGRVTADGKHIQKFSGGGSDLKENEEYEIPEKARIKYKELLRVYNNNPQAVPESVFQSRVARENVEKLKSGDLVYFKHNEKYVEDIVPVRISRTVDDRMIGKRMSADLRPCHGDWVEDGDLSALNAYPEKRLLLRHPKGLCPACRLFGTGSYKGRVRFGFASLENDPEWLIPGKNPGDPFHGGPVMLSLLERPRPTWSIPGSDNKFKVPGRKFYVHHHAWKTIKDGNHPTTGKAIEQSPNNRTVEALAGGNSFSFEIAFENLKEWELGLLIHSLQLEKGLAHKLGMAKSMGFGSVEIDVESVRLRKDWKQWRNGNSEIPNWLGKGFAKLKEWFRDELDFIENLKKLLWFPEGDQAPRVCYPMLRKKDDPNGNSGYEELKDGEFKKEDRQKKLTTPW TPWA SEQ IDMTTTMKISIEFLEPFRMTKWQESTRRNKNNKEFVRGQAFARWHRNKK NO: 4DNTKGRPYITGTLLRSAVIRSAENLLTLSDGKISEKTCCPGKFDTEDKD huDiCas7-RLLQLRQRSTLRWTDKNPCPDNAETYCPFCELLGRSGNDGKKAEKKD 11-WRFRIHFGNLSLPGKPDFDGPKAIGSQRVLNRVDFKSGKAHDFFKAYE R1045-VDHTRFPRFEGEITIDNKVSAEARKLLCDSLKFTDRLCGALCVIRFDEY GGGS-TPAADSGKQTENVQAEPNANLAEKTAEQIISILDDNKKTEYTRLLADAI R1122RSLRRSSKLVAGLPKDHDGKDDHYLWDIGKKKKDENSVTIRQILTTSADTKELKNAGKWREFCEKLGEALYLKSKDMSGGLKITRRILGDAEFHGKPDRLEKSRSVSIGSVLKETVVCGELVAKTPFFFGAIDEDAKQTDLQVLLTPDNKYRLPRSAVRGILRRDLQTYFDSPCNAELGGRPCMCKTCRIMRGITVMDARSEYNAPPEIRHRTRINPFTGTVAEGALFNMEVAPEGIVFPFQLRYRGSEDGLPDALKTVLKWWAEGQAFMSGAASTGKGRFRMENAKYETLDLSDENQRNDYLKNWGWRDEKGLEELKKRLNSGLPEPGNYRDPKWHEINVSIEMASPFINGDPIRAAVDKRGTDVVTFVKYKAEGEEAKPVCAYKAESFRGVIRSAVARIHMEDGVPLTELTHSDCECLLCQIFGSEYEAGKIRFEDLVFESDPEPVTFDHVAIDRFTGGAADKKKFDDSPLPGSPARPLMLKGSFWIRRDVLEDEEYCKALGKALADVNNGLYPLGGKSAIGYGQVKSLGIKGDDKRISRLMNPAFDETDVAVPEKPKTDAEVRIEAEKVYYPHYFVEPHKKVEREEKPCGHQKFHEGRLTGKIRCKLITKTPLIVPDTSNDDFFRPADKEARKEKDEYHKSYAFFRLHKQIMIPGSELRGMVSSVYETVTNSCFRIFDETKRLSWRMDADHQNVLQDFLPGRVTADGKHIQKFSETARVPFYDKTQKHFDILDEQEIAGEKPVRMWVKRFIKRGGGSRDSRYQKAFQEIPENDPDGWECKEGYLHVVGPSKVEFSDKKGDVINNFQGTLPSVPNDWKTIRTNDFKNRKRKNEPVFCCEDDKGNYYTMAKYCETFFFDLKENEEYEIPEKARIKYKELLRVYNNNPQAVPESVFQSRVARENVEKLKSGDLVYFKHNEKYVEDIVPVRISRTVDDRMIGKRMSADLRPCHGDWVEDGDLSALNAYPEKRLLLRHPKGLCPACRLFGTGSYKGRVRFGFASLENDPEWLIPGKNPGDPFHGGPVMLSLLERPRPTWSIPGSDNKFKVPGRKFYVHHHAWKTIKDGNHPTTGKAIEQSPNNRTVEALAGGNSFSFEIAFENLKEWELGLLIHSLQLEKGLAHKLGMAKSMGFGSVEIDVESVRLRKDWKQWRNGNSEIPNWLGKGFAKLKEWFRDELDFIENLKKLLWFPEGDQAPRVCYPMLRKKDDPNGNSGYEELKDGEFKKEDRQKKLTTPWTPW A

Table 4 below shows examples of target ssRNA sequences (5′-3′).

TABLE 4 SEQ ID NO/Description Sequence SEQ ID NO: 5ggAUUACCCAUGUCGAAGACAACAAAG tgRNA for structural analysis SEQ ID NO: 6/5Cy5/GGGGUUGGAAAGCCGGUUUUCUUUGAUG 60-nt processing targetUCACGGAACCUUUGUUGUCUUCGACAUGGGUA SEQ ID NO: 7/5Cy5/CCGGCCAUUCAAACAUGAGGAUUACCCAU 60-nt MS2 ssRNA targetGUCGAAGACAACAAAGAAGUUCAACUCUUUA

Table 5 below shows examples of Cas7-11 guide sequences (5′-3′).

TABLE 5 SEQ ID NO/Description Sequence SEQ ID NO: 8ggUGAUGUCACGGAACCUUUGUUGUCUUCGACA crRNA for structural analysis UGGGUAAUSEQ ID NO: 9 GGUUGAUGUCACGGAACCUUUGUUGUCUUCGA Mature crRNA for MS2CAUGGGUAAUCCUCAU ssRNA target SEQ ID NO: 10GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGACAUGGGUAAUCC ssRNA target, WT UCAU SEQ ID NO: 11GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACGUUUGUUGUCUUCGACAUGGGUAAUCC ssRNA target, mismatch 1 UCAUSEQ ID NO: 12 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCAUUGUUGUCUUCGACAUGGGUAAUCC ssRNA target, mismatch 2 UCAUSEQ ID NO: 13 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUAUGUUGUCUUCGACAUGGGUAAUCC ssRNA target, mismatch 3 UCAUSEQ ID NO: 14 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUAGUUGUCUUCGACAUGGGUAAUCC ssRNA target, mismatch 4 UCAUSEQ ID NO: 15 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUCUUGUCUUCGACAUGGGUAAUCC ssRNA target, mismatch 5 UCAUSEQ ID NO: 16 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGAUGUCUUCGACAUGGGUAAUCC ssRNA target, mismatch 6 UCAUSEQ ID NO: 17 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUAGUCUUCGACAUGGGUAAUCC ssRNA target, mismatch 7 UCAUSEQ ID NO: 18 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUCUCUUCGACAUGGGUAAUCC ssRNA target, mismatch 8 UCAUSEQ ID NO: 19 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGACUUCGACAUGGGUAAUCC ssRNA target, mismatch 9 UCAUSEQ ID NO: 20 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUGUUCGACAUGGGUAAUCC ssRNA target, mismatch 10 UCAUSEQ ID NO: 21 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCAUCGACAUGGGUAAUCC ssRNA target, mismatch 11 UCAUSEQ ID NO: 22 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUACGACAUGGGUAAUCC ssRNA target, mismatch 12 UCAUSEQ ID NO: 23 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUGGACAUGGGUAAUCC ssRNA target, mismatch 13 UCAUSEQ ID NO: 24 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCCACAUGGGUAAUCC ssRNA target, mismatch 14 UCAUSEQ ID NO: 25 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGUCAUGGGUAAUCC ssRNA target, mismatch 15 UCAUSEQ ID NO: 26 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGAGAUGGGUAAUCC ssRNA target, mismatch 16 UCAUSEQ ID NO: 27 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGACUUGGGUAAUCC ssRNA target, mismatch 17 UCAUSEQ ID NO: 28 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGACAAGGGUAAUCC ssRNA target, mismatch 18 UCAUSEQ ID NO: 29 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGACAUCGGUAAUCC ssRNA target, mismatch 19 UCAUSEQ ID NO: 30 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGACAUGCGUAAUCC ssRNA target, mismatch 20 UCAUSEQ ID NO: 31 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGACAUGGCUAAUCC ssRNA target, mismatch 21 UCAUSEQ ID NO: 32 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGACAUGGGAAAUCC ssRNA target, mismatch 22 UCAUSEQ ID NO: 33 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUUUGUUGUCUUCGACAUGGGUUAUCC ssRNA target, mismatch 23 UCAUSEQ ID NO: 34 GGGGUUGGAAAGCCGGUUUUCUUUGAUGUCAC Pre-mature crRNA for MS2GGAACCUAUGUUGACUUCGACAUGGGUAAUCC ssRNA target, mismatch 3/9 UCAUSEQ ID NO: 35 GUUGAUGUCACGGAACUGCAGCCAGCUUUCCG Gluc targeting guide, WTGGCAUUGGCUUCCAU SEQ ID NO: 36 GUUGAUGUCACGGAACGGUAAUGCCUGGCUUGNon-targeting guide UCGACGCAUAGUCUG SEQ ID NO: 37GUUGAUGUCACGGAACCAGUGUUGGUAGGAGU PPIB targeting guide UUGUUACAAAAGUGASEQ ID NO: 38 GUUGAUGUCACGGAACGGUUAUAGCUUGACAA MALAT1 targeting guideGCAAUUAACUUUAAA

Example 1 Protein and RNA Preparation

A gene encoding D. ishimotonii Cas7-11 (residues 1-1601) was amplifiedby PCR and cloned into the modified pET vector (Novagen), in whichCas7-11 has an N-terminal maltose-binding protein (MBP) and a C-terminalHis₆-tag. The two inactivating mutations (D429A/D654A) were introducedinto Cas7-11 by a PCR-based method, and the sequence was confirmed byDNA sequencing. The MBP-Cas7-11 (D429A/D654A)-His₆ protein was expressedin Escherichia coli Rosetta2 (DE3) (Novagen) by inducing with 0.1 mMisopropyl β-D-thiogalactopyranoside (Nacalai Tesque) at 20° C.overnight. The E. coli cells were lysed by sonication and the lysate wasclarified by centrifugation. The supernatant was applied to Ni-NTAsuperflow (QIAGEN) and the MBP-Cas7-11 (D429A/D654A)-His₆ protein waseluted by buffer A (20 mM Tris-HCl, pH 8.0, 20 mM imidazole, 1 M NaCl, 3mM 2-mercaptoethanol, and 1 mM phenylmethylsulfonyl fluoride) with 300mM imidazole. The protein was further purified by chromatography onAmyrose resin (NEB), HiTrap Heparin (GE Healthcare), and HiLoad 16/600Superdex 200 (GE Healthcare) columns. The crRNA (39 nucleotides plus 5′GG for in vitro transcription) and target RNA (25 nucleotides plus 5′ GGfor in vitro transcription) were transcribed in vitro with T7 RNApolymerase and purified by 10% denaturing (7 M urea) polyacrylamide gelelectrophoresis. The purified materials were stored at −80° C. untiluse.

Example 2 Cryo-EM Grid Preparation and Data Collection

A Cas7-11-crRNA-target RNA complex was reconstituted by mixing thepurified MBP-Cas7-11 protein, the 39-nucleotide crRNA, and the25-nucleotide target RNA, at a molar ratio of 1:1.2:1.5. The complex waspurified by size-exclusion chromatography on a Superose6 Increase 10/300column (GE Healthcare), equilibrated with the buffer containing 20 mMHepes-NaOH, pH 7.0, 150 mM NaCl, 2 mM MgCl₂, and 1 mM DTT. The peakfraction containing Cas7-11-:crRNA-target RNA complex was concentratedto 1.5 A260 units using an Amicon Ultra-4 filter (10 kDamolecular-weight cutoff; Millipore). The samples (3 μl) were thenapplied to freshly glow-discharged Au 300 mesh R1.2/1.3 grids(Quantifoil) in a Vitrobot Mark IV (FEI) at 4° C. with a waiting time of10 sec and a blotting time of 4 sec under 100% humidity conditions. Thegrids were plunge-frozen into liquid ethane cooled at liquid nitrogentemperature.

The cryo-EM data were collected using a Titan Krios G3i microscope(Thermo Fisher Scientific), running at 300 kV and equipped with a GatanQuantum-LS Energy Filter (GIF) and a Gatan K3 Summit direct electrondetector. Micrographs were recorded at a nominal magnification of×105,000 with a pixel size of 0.83 Å in a total exposure of 52 e⁻/A² per64 frames by the correlated double sampling mode. The data wereautomatically acquired by the image shift method using the SerialEMsoftware (Mastronarde, 2005), with a defocus range of −0.8 to −1.6 μm,and 2,781 movies were acquired.

Example 3 Image Processing

Data processing was performed using a combination of cryoSPARC v3.2.0and Relion3.1 software packages. The dose-fractionated movies werealigned using the Patch motion correction and the contrast transferfunction (CTF) parameters were estimated using Patch-Based CTFestimation in cryoSPARC. Particles were automatically picked using Blobpicker in cryoSPARC followed by multiple times of reference free 2Dclassification to curate particle sets. The particles were furthercurated by cryoSPARC Heterogeneous Refinement (N=6) using the mapderived from cryoSPARC Ab initio Reconstruction as a template. The bestclass containing 581,179 particles was refined using Homogeneousrefinement followed by non-uniform refinement, yielding a map at 2.46 Åresolution. To improve the quality of 3D reconstruction in the legdomain, the particles were imported into Relion and subjected to 3Dclassification without alignment using a mask for the leg domain. Theselected 135,109 particles were imported back to cryoSPARC andnon-uniform refinement after local motion correction yielded a map at2.45 Å resolution, according to the Fourier shell correlation(FSC)=0.143 criterion. The local resolution was estimated by BlocRes incryoSPARC.

Example 4 Model Building and Validation

A model was built using Nautilus and Buccaneer in CCP-EM package andmanually built using COOT against the density map sharpened usingDeepEMhancer. The model was refined using Real-space refinement inPHENIX with the secondary structure restraints. The structure validationwas performed using MolProbity from the PHENIX package. The curverepresenting model vs. full map was calculated using phenix.mtriage,based on the final model and the full, filtered, and sharpened map. Thestatistics of the 3D reconstruction and model refinement are summarizedin Table 6. The cryo-EM density maps were calculated with UCSF ChimeraX,and molecular graphics figures were prepared with CueMol(http://www.cuemol.org).

TABLE 6 Data Collection and Structural Refinement Data collection andprocessing Magnification 105,000 Voltage (kV) 300 Electron exposure(e⁻/Å²) 76.5/72.8 Defocus range (μm) −0.6 to −1.5 Pixel size (Å) 0.83Initial particle images (no.) 4,452,943 Refinement PDB ID 7WAH EMDB IDEMD-32385 Final particle images (no.) 135,109 Map resolution (Å) 2.45FSC threshold 0.143 Model resolution (Å) 2.89 FSC threshold 0.5 Mapsharpening B factor (Å²) −46.8 Model composition Non-hydrogen atoms13,228 Protein residues 1,473 RNA nucleotides 62 Ligands 4 R.m.s.deviations Bond lengths (Å) 0.002 Bond angles (°) 0.507 ValidationClashscore 8.27 Poor rotamers (%) 0.47 Ramachandran plot (%) Favored97.32 Allowed 2.68 Outliers 0.00

Example 5 In Vitro Pre-crRNA and Target RNA Cleavage Assays

In vitro Pre-crRNA and target RNA cleavage assays were performed (FIGS.15-19 ). 5′-labelled ssRNA processing and cleavage targets weresynthesized by Integrated DNA Technologies. crRNA and unlabeled ssRNAtargets were prepared with the HiScribe™ T7 High Yield RNA Synthesis Kit(New England Biolabs). In vitro transcription products were thenpurified with either RNAClean XP magnetic beads (Beckman Coulter) or RNAClean and Concentrator columns (R1017, Zymo Research). In vitro cleavageassays were performed with 233 nM purified Cas7-11, 30 nM of5′-Cy5-labelled ssRNA targets and 200 nM crRNA in nuclease assay buffer(40 mM Tris-HCl, pH 7.5, 60 mM NaCl and 6 mM MgCl₂) supplemented with 4U of RNase inhibitor, murine (M0314S, New England Biolabs). Forpre-crRNA processing reactions, crRNA was omitted, and pre-crRNA wasused in place of the labelled ssRNA target. Reactions were incubated for1 h at 37° C. (unless otherwise indicated) and then quenched withaddition of proteinase K, EDTA and urea (final concentrations 1 mg ml⁻¹proteinase K, 6 mM EDTA and 400 μM urea) for 30 min at 50° C. Reactionswere denatured with 4.5 M urea denaturing buffer at 95° C. for 5 min,and loaded onto a 10% (for pre-crRNA processing) or 6% (for target ssRNAcleavage) Novex PAGE Tris-borate-EDTA (TBE)-urea gel (EC6885BOX,Invitrogen), which was run at 200 V for 35 min at 60° C. Gels wereimaged using an Odyssey scanner (LI-COR Biosciences).

Example 6 Mammalian Cell Culture

Mammalian experiments were performed using the HEK293FT cell line,acquired from and authenticated by American Type Culture Collection(ATCC). HEK293FT cells were grown in Dulbecco's modified Eagle mediumwith high glucose, sodium pyruvate and GlutaMAX (Thermo FisherScientific), additionally supplemented with 1× penicillin-streptomycin(Thermo Fisher Scientific) and 10% fetal bovine serum (Thermo FisherScientific) and passaged using TrypLE Express (Thermo FisherScientific). Cells were maintained at 37° C. and 5% CO₂. Fortransfection of HEK293FT cells, cells were plated 16 h beforetransfection at seeding densities of 1.5×10⁴ cells per well in a 96-wellplate or 1.5×10⁶ cells per T25 flask, allowing cells to reach 90%confluency by transfection. Cells were then transfected withLipofectamine 3000 (Thermo Fisher Scientific) following themanufacturer's protocol with 200 ng total plasmid per well in a 96-wellplate and 7.7 μg total plasmid in a T25 flask.

Example 7 RNA Knockdown Assay in Human Cells

To assess RNA knockdown in mammalian cells with reporter constructs, 80ng of the DiCas7-11 expression vector was co-transfected with 80 ng ofguide expression plasmid and 40 ng of the dual luciferase reporter.After 48 h, the medium containing the secreted luciferase was collectedand luciferase activity was measured using the Gaussia Luciferase Assayreagent (GAR-2B; Targeting Systems) and Cypridina (Vargula) luciferaseassay reagent (VLAR-2; Targeting Systems) kits. Assays were performed inwhite 96-well plates on a plate reader (Biotek Synergy Neo 2) with aninjection protocol. All replicates performed were biological replicates.Luciferase measurements were normalized by dividing the Gluc values bythe Cluc values, thus normalizing for any variation between wells.

For targeting of endogenous genes, 100 ng of the DiCas7-11 expressionvector was co-transfected with 100 ng of guide expression plasmid. After48 h, the cells were lysed and the RNA were collected using a previouslydescribed method (Julia Joung, Nat. Protocol) with the Revertaid RTReverse Transcription Kit (Thermo Fisher), following the manufacturer'sprotocol. Using Fast Advanced Master Mix (Thermo Fisher Scientific),gene expression was measured using the cDNA by TaqMan qPCR probes forthe KRAS, PPIB, and MALAT1 transcripts (Thermo Fisher Scientific) aswell as the GAPDH control probe (Thermo Fisher Scientific). qPCRreactions were read out on a Bio-Rad CFX384 Touch Real-Time PCRDetection System, with three 10-μl technical replicates in 384-wellformat. No statistical methods were used for determining sample size. Noblinding or randomization methods were used during these experiments.

Example 8 Truncated DiCas7-11 AAV Production and Delivery

AAV was prepared by designing vectors with truncated Cas7-11 expressionusing an EFS promoter or guide expression with a U6 or tRNA promoter.HEK293FT cells were transfected in T25 flasks using Lipofectamine 3000(Thermo Fisher Scientific) with 2.0 μg of cargo plasmid, 1.8 μg AAV8capsid vector and 3.9 μg AAV helper pAdDeltaF6 plasmid (Addgene 112867)per T25 flask according to the manufacturer's protocol. Two days afterthe transfection, the medium containing the loaded viral vector wasfiltered using a 0.45-μm filter (Sigma Aldrich), concentrated by anAmicon Ultra-15 Centrifugal Filter Unit (MWCO 100 kDa), washed once with1×DPBS (Thermo Fisher), and the final product was stored at −80° C. Toevaluate knockdown on the luciferase reporter expressed in HEK293FTcells, AAV was added at varying titres to the 40,000 cells per well in a96-well plate by spinfection at 2,000 g and 37° C. for 2 h. The dualluciferase reporter plasmid was subsequently transfected to the HEK293FTcells at 100 ng per well with Lipofectamine 3000. Cell media washarvested for luciferase chemiluminescence measurement 48 h later. AAVgenome titer was determined by RT-qPCR, using a pair of primerstargeting the EFS promoter in the Fast SYBR Green Master Mix (AppliedBiosystems).

Example 9 Structure of Cas7-11 Effector Complex

The cryo-EM structure of D. ishimotonii Cas7-11, catalyticallyinactivated by D429A/D654A mutations, in complex with a 39-nt crRNA(U(−14)-U25) and its complementary 25-nt target RNA (G1*-A25*) at 2.5-Aresolution were determined (FIGS. 1A, 1B, and 8A-8D).

Cas7-11 consists of four Cas7 domains (Cas7.1-Cas7.4), and a Cas11domain, interspaced with four interdomain linkers (L1-L4), with Cas7.4harboring an additional large insertion (INS)(residues 979-1293) domainand a C-terminal extension (CTE)(residues 1507-1601) domain (FIGS. 1Cand 1D). Cas7-11 has a baby-like structure, with Cas7.1,Cas7.2-Cas7.4/CTE, INS, and Cas11 corresponding to the head, body, legs,and arms, respectively. The Cas7.1-Cas7.4 domains stack and form aright-handed helical filament. Cas11 interacts with Cas7.2 and Cas7.3 atthe midpoint and is directly connected with Cas7.1 and Cas7.2 by L1(residues 238-259) and L2 (residues 365-401) which, are disordered inthe present structure, indicating their flexibility. The repeat-derivedregion (referred to as the 5′ tag) of the crRNA is anchored by Cas7.1and Cas7.2, while the duplex formed by the spacer-derived region of thecrRNA and target RNA is recognized by Cas7.2-Cas7.4 and INS. CTEextensively interacts with Cas7.3, Cas7.4, and L4, structurallyreinforcing the Cas7-11 architecture.

Example 10 Domain Structure of Cas7-11

The domain structure of Cas7-11 effector was assessed. The Cas7.1-Cas7.4domains contain a modified RRM (RNA recognition motif) fold (also knownas a ferredoxin-like fold), consisting of a four-stranded antiparallelβ-sheet flanked by two α helices in a βαββαβ topology, as commonlyobserved in the type III-A/B Cas7 proteins (Csm3/Cmr4) (Taylor et al.2015; Osawa et al. 2015; You et al. 2019; and Jia et al. 2019,incorporated herewith in their entirety by reference) (FIGS. 2A-2E and1A). Unexpectedly, unlike Csm3 and Cmr4, Cas7.1-Cas7.4 contain a zincfinger motif between the α1 helix and β2 strand in the RRM fold, withzinc ions in each of Cas7.1-Cas7.4 coordinated by C86/C115/C123/C126,C463/C472/C474/C477, H703/C706/C708/C711, and C965/C1312/C1342/C1345,respectively (FIGS. 2A-2D). These zinc-coordinating residues are highlyconserved among the Cas7-11 orthologs (FIGS. 10A-10C), indicating thatzinc fingers are shared structural features of the Cas7-11 proteins.These residues are not conserved in the Cas7.1 and Cas7.3 subunits ofthe Cas7×3 family (homologous to the Cas7.2 and Cas7.4 domains ofCas7-11, respectively), but are present in a subset of the Cas7.2subunit of Cas7×3, implying the zinc finger insertion succeeded thefusion event that produced the Cas7×3 family. Cas7.1-Cas7.3 have theirRRM fold with a thumb-like β-hairpin between the β2 and β3 strands, asobserved in Csm3 and Cmr4 (FIGS. 2A-2C and 9A). Cas7.1-Cas7.3 alsocontain unique structural elements, consistent with their distinctfunctional roles, such as pre-crRNA processing and target RNA cleavage.Cas7.1 and Cas7.2/Cas7.3 possess the catalytic residues for pre-crRNAprocessing (H43) and target RNA cleavage (D429 and D654) between the β1strand and the α1 helix, respectively. Unlike Cas7.1 and Cas7.2, Cas7.3has an additional β-hairpin between the β1strand and the α1 helix (FIG.2C). In line with its later fusion into the single effector during theevolution of Cas7-11, Cas7.4 adopts the RRM fold highly divergent fromthose of Cas7.1-Cas7.3 and contains a larger insertion (residues1365-1452), rather than a thumb-like β-hairpin, between the β2 and β3strands (FIG. 2D). In addition, the INS domain (residues 966-1311) isinserted within the zinc finger motif of Cas7.4. The Cas11 domain adoptsa five-helix bundle similar to the other type III Cas11 proteins (Csm2and Cmr5) (FIGS. 2E and 9B).

The INS domain comprises two five-stranded β-barrels and additionalstructural elements, including an a helix and a four-strandedantiparallel β-sheet (FIGS. 2F and 9C). Computational structuralcomparison revealed that the two β-barrels of INS are structurallysimilar to cold shock proteins, such as CspB, and the rest of INS lacksstructural similarity with any other known proteins (e.g., Lisa Holm,“Using Dali for Protein Structure Comparison,” Methods Mol. Biol. 2020;2112:29-42, doi 10.1007/978-1-0716-0270-6_3; and Schindelin, H.,Marahiel, M. & Heinemann, U., “Universal nucleic acid-binding domainrevealed by crystal structure of the B. subtilis major cold-shockprotein,” Nature 364, 164-168 (1993), doi 10.1038/364164a0, incorporatedherewith in their entirety by reference). The INS domain is lessresolved in the density map relative to the other domains (FIG. 8D),suggesting its flexibility, consistent with limited contacts between theINS domain and the rest of the Cas7-11 domains.

Example 11 Domain Interfaces of Cas7-11

The domain interfaces of Cas7-11 were assessed. Cas7.1-Cas7.4 form acentral filament in the Cas7-11 structure (FIGS. 11A and 11B).Cas7.1-Cas7.3 commonly employ two interfaces for the interaction withtheir adjacent Cas7 domains (Cas7.2-Cas7.4), as in the Cas7 filaments(Csm3/Cmr4) in the type III-A/B effectors (Taylor et al. 2015; Osawa etal. 2015; You et al. 2019; and Jia et al. 2019, incorporated herewith intheir entirety by reference). At the primary interface, the β1-α1 andα2-β4 regions of a Cas7 domain interact with the β3 strand of theadjacent Cas7 domain. At the second interface, the thumb-like β-hairpin(in the β2-β3 region) of a Cas7 domain contacts the α1 helix of theadjacent Cas7 domain (FIGS. 11A and 11B). In addition to the twointerfaces, each Cas7 domain forms distinct interactions with theiradjacent Cas7 domains, consistent with the structural variations amongthe Cas7.1-Cas7.4 domains. The thumb-like β-hairpins of Cas7.1 andCas7.2 form additional contacts with the zinc-finger loops (in the α1-β2region) of Cas7.2/Cas7.3 and Cas7.3/Cas7.4, respectively (FIGS. 11A and11B). In addition, the thumb-like β-hairpins of Cas7.2 and Cas7.3interact with Cas7.3 (the additional β-hairpin) and Cas7.4 (thezinc-finger loop and the β1-α1 and β2-β3 regions), respectively (FIGS.11A and 11B). The L3 and L4 linkers also contribute to stabilizing theCas7.1-Cas7.4 filament structure. The L3 linker reinforces the interfacebetween Cas7.1 and Cas7.2, while the L4 linker adopts a V-shapedconformation and extensively interacts with Cas7.2, Cas7.3, Cas7.4, andCTE (FIGS. 11C and 11D). Cas11 mainly interacts with the β1-α1 regionsof Cas7.2 and Cas7.3 (FIGS. 11A and 11B).

Example 12 Pre-crRNA Processing Mechanism

The Pre-crRNA processing mechanism was assessed. The 5′ tag region(U(−14)-C(−1)) of the crRNA adopts a single-stranded conformation and isextensively recognized by Cas7.1 and Cas7.2 (FIGS. 3A, 3B, 4A, and 12 ).The thumb-like β-hairpin of Cas7.1 intercalates between A(−2) and G(−4),resulting in the base flipping of A(−3) (FIG. 4B). The nucleobases ofA(−2) and G(−4) stack with F186N172 and F187 in the β-hairpin,respectively, while the nucleobases of C(−1) and G(−4)hydrogen bond withA183 and R35, respectively. A(−3) stacks with V176, I450 and R453 inCas7.2. C(−6) is also flipped out, and A(−7), G(−5), and C(−8) form atriple stack, which is sandwiched by R35 and P471 (FIGS. 4C and 4D). Theflipped-out C(−6) forms a stacking interaction with R444 and T484, andmultiple hydrogen bonds with E13, R448, and V485 (FIG. 4C), consistentwith a previous finding that the mutation of C(−6) completely inhibitedtarget RNA cleavage (Ozcan et al. 2021, incorporated herewith in itsentirety by reference). G(−5) adopts the syn conformation, and hydrogenbonds with Q103 and the A(−7) backbone phosphate (FIG. 4D). Thenucleobases of U(−9), G(−10), U(−11), and A(−12) hydrogen bond with Q37,T92/R102, D91, and T59, respectively (FIG. 4E). U(−14), the firstnucleotide of the 5′ tag, forms hydrogen-bonding and stackinginteractions with T59/H149/F150 and N152, respectively (FIG. 4E). Inaddition to these base-specific contacts, the 5′ tag region formssequence-independent backbone interactions with Cas7.1 and Cas7.2 (FIG.3B). The 14-nt 5′ tag sequence is highly conserved among Cas7-11orthologs (Ozcan et al. 2021; and van Beljouw et al. 2021, incorporatedherewith in their entirety by reference), therefore the 5′ tag regionsmay be recognized by Cas7-11 orthologs in a similar manner.

D. ishimotonii Cas7-11 cleaves pre-crRNAs between U(−15) and U(−14) ofthe direct repeat sequence in a metal-independent manner, to producemature crRNAs with a 14-nt 5′ tag sequence (Ozcan et al. 2021,incorporated herewith in its entirety by reference). Metal-independentRNA hydrolysis via acid-base catalysis yields a 3′ product with a5′-hydroxy group and a 5′ product with a 2′,3′-cyclic phosphate group,which is then converted to a 3′ phosphate group (Yang 2011, incorporatedherewith in its entirety by reference). As the crRNA used for the cryoEManalysis contains G(−15), rather than U(−15), for an in vitrotranscription reaction, a density corresponding to aguanosine-3′,5′-diphosphate (pGp) adjacent to U(−14), which contains a5′-hydroxy group was observed (FIG. 4F). These structural observationsindicates that, during the complex reconstitution, Cas7-11 cleaves thephosphodiester bond between G(−15) and U(−14) of the crRNA via anacid-base catalytic mechanism. The pGp molecule is surrounded byresidues R41, H43, R53, Y55, N152, and K158 in the Cas7.1 domain (FIG.4F), and H43, Y55, and N152 are highly conserved in the Cas7-11orthologs (FIGS. 10A-10C). Thus, the H43A, Y55A, and N152A Cas7-11mutants were prepared and their pre-crRNA processing and target RNAcleavage activity in vitro were tested (FIG. 14 ). It was observed thatthe Y55A and N152A mutants exhibit reduced pre-crRNA processingactivities (FIG. 4G), confirming the functional importance of Y55 andH152 in the pre-crRNA processing. Notably, the H43A mutant lacks theprocessing activity (FIG. 4G), indicating that H43 is critical for thepre-crRNA processing. Thus, H43 may serve as a general base anddeprotonates the 2′-hydroxy group of U(−15), which then nucleophilicallyattacks on the scissile phosphate between U(−15) and U(−14). Thesemutants were capable of target RNA cleavage (FIG. 4H). Therefore,Cas7-11 processes its pre-crRNAs in the Cas7.1 domain via an acid-basecatalytic mechanism.

Example 13 Target RNA Recognition Mechanism

The RNA recognition mechanism was assessed. The spacer region (C1-A23,except for U4 and C10) of the crRNA base pairs with the target RNA toform the guide-target duplex, assisted by the INS domain (FIGS. 3A, 3B,5A, 5B, and 12 ). The last base pair, A23-U23*, is capped by R1125 inthe INS domain, while A24 is splayed out from the duplex andaccommodated within a pocket formed by R1045, H1098, and K1099 in theINS domain (FIGS. 5A and 5B). U25 in the crRNA and U24*/A24* in thetarget RNA are disordered in the structure. These structuralobservations indicate that the 23-nt spacer sequence serves as a guidesegment, consistent with a previous study showing that 22-25-nt, but not19-nt, spacers support Cas7-11-mediated target RNA cleavage (Ozcan etal. 2021, incorporated herewith in its entirety by reference).

The 23-bp guide-target duplex consists of six segments (segments 1-6),consisting of successive base pairs, with the flipped-out nucleotides atfourth and tenth positions and kinks at the 13th-14th, 15th-16th, and19th-20th base pairs (FIGS. 3A and 12 ). The thumb-like β-hairpins inCas7.2 and Cas7.3 intercalate within the guide-target duplex, flippingthe fourth and tenth nucleotides, respectively (FIGS. 5C and 5D).Accordingly, the segments 1-3 (C1-G1*-C13-G13*) adopt an underwoundribbon-like conformation resembling a ladder, rather than a doublehelix, similar to the guide-target duplexes in the type III-A/B effectorcomplexes (Taylor et al. 2015; Osawa et al. 2015; You et al. 2019; andJia et al. 2019, incorporated herewith in their entirety byreference)(FIGS. 5C, 5D, and 13A-13C). The U3-A3* and G5-C5* base pairsstack with F517 and L516 in the β-hairpin in Cas7.2, respectively, whilethe U9-A9* and U11-A11* base pairs stack with F757 and K756 in theβ-hairpin in Cas7.3, respectively (FIG. 5D). The flipped-out U4 and C10nucleobases are sandwiched by I504/V682 of Cas7.3 and M953/K1489 ofCas7.4, respectively. Appropriate for their involvement in recognition,three of these residues (I504, M953, and K1489) are conserved in boththe Cas7-11 and Cas7×3 families, with V682 conserved within the Cas7-11clade. The β2-β3 loop of Cas7.4 (equivalent to the thumb-like β-hairpinsof Cas7.2 and Cas7.3) penetrates between the C13-G13* and G14-C14* basepairs in the guide-target duplex, resulting in a kink between thesegments 3 and 4 (FIGS. 5A and 5B). The C13-G13* and G14-C14* base pairsinteract with L1391 and E1392 in the β2-β3 loop of Cas7.4, respectively.L1564 in CTE and A981/I1292 in INS are wedged into the 15th-16th and19th-20th base pairs, thereby forming a kink between the segments 4 and5, and 5 and 6, respectively (FIG. 5B). Moreover, the sugar-phosphatebackbone of the crRNA spacer region is extensively recognized by theCas7.2-Cas7.4, INS, and CTE domains (FIG. 3B). These structuralobservations agree with conservation across these nuclease families andexplain the mechanism of crRNA-guided target RNA recognition by Cas7-11.

Example 14 Target RNA Cleavage Mechanism

The RNA cleave mechanism was assessed D. ishimotonii Cas7-11 cleaves thetarget RNA with conserved aspartate residues (D429 and D654), generatingtwo cleavage sites separated by 5-6 nt near the 3′ end of thespacer-complementary region, although the precise cleavage sites werenot determined (Ozcan et al. 2021, incorporated herewith in its entiretyby reference). As for the Cas7-11 (D429A/D654A)structure, A429 in Cas7.2and A654 in Cas7.3 are located close to the phosphodiester bonds betweenA3* and A4* and between U9* and C10* in the target RNA, respectively(FIGS. 5C and 5D), appropriate for their role as the catalytic residuesfor target RNA cleavage. In addition, the backbone phosphate groupsbetween A3* and A4* and between U9* and C10* are recognized by Y360 andH306 in Cas11, respectively (FIG. 5D). The configuration of D429 andD654 of Cas7-11 relative to the target RNA is similar to that of thecatalytic aspartate residues (D33 in Csm3) of the type III-A effectorcomplex (FIGS. 13A-13C). These structural observations indicates that D.ishimotonii Cas7-11 cleaves a target RNA between the third and fourthnucleotides (site 1) and between the ninth and tenth nucleotides (site2), using D429 in Cas7.2 and D654 in Cas7.3, respectively, consistentwith Scalindua brodae Cas7-11 (Sb-gRAMP) that cleaves a target RNA atthese two positions (van Beljouw et al. 2021, incorporated herewith inits entirety by reference).

In vitro cleavage activities of Cas7-11 against a 5′-fluorescentlylabelled, 60-nt ssRNA target, using crRNAs with mismatched spacers(mm1-23) were measured to validate the structural observations. Mm4 andmm10 were observed to do not affect the target RNA cleavage (FIG. 5E),consistent with the base flipping at positions 4 and 10 in the Cas7-11structure. In contrast, mm3 and mm9 was observed to abolish the cleavageat the sites 1 and 2 on the target RNA (FIG. 5E), respectively,indicating that base pairing at positions 3 and 9 are critical for theRNA target cleavage at the sites 1 and 2, respectively. Accordingly,double mismatch (mm3/9) abolishes the target RNA cleavage (FIG. 5E).While mm1 and mm2 attenuates the cleavage at the site 1 on the targetRNA, mismatches at the other positions (mm5-8 and mm11-23) do notsubstantially affect the RNA cleavage (FIG. 5E). Overall, the structuraland functional data shows that Cas7-11 cleaves ssRNA targets after thirdand ninth nucleotides, counting from the 3′ end of thespacer-complementary region, by the Cas7.2 and Cas7.3 domains,respectively.

Example 15 Structure-guided Engineering of Compact Cas7-11 Variants

The structure-guided engineering of compact Cas7-11 variants wasassessed. Three Cas7-11 deletion variants, ΔINS-1 (1,219 aa), ΔINS-2(1,392 aa), and ΔINS-3 (1,530 aa), in which residues 979-1293,1007-1220, and 1046-1121 are deleted and replaced with a GGGS linker,respectively were engineered (FIG. 6A). Their target RNA cleavageactivities in vitro were measured, and it was found that the truncatedCas7-11 variants exhibit the same level of in vitro cleavage activity onthe target RNA as compared to the full-length Cas7-11 (FIG. 6B). Using adual-luciferase assay (Özcan et al. 2021, incorporated herewith in itsentirety by reference), these truncated ΔINS Cas7-11 variants weredemonstrated to preserve cleavage activity as compared to the wild-type(WT) Cas7-11 in mammalian cells (FIG. 6C). In contrast, the ΔCas11variant (A259-GGGS-G398) greatly decreases Gluc transcript knockdown(FIG. 6C), indicating the importance of the Cas11 domain for target RNAcleavage. Both the WT Cas7-11 and ΔINS Cas7-11 variants also effectivelyknocks down endogenous mRNA targets (PPIB and MALAT) with comparableefficiency (FIG. 6D). Given its activity in mammalian cells and value asa tool for in vivo RNA-targeting, the smallest functional Cas7-11truncation, ΔINS-1, was named as Cas7-11S. The reduced size of Cas7-11Sand the ΔINS variants compared to WT Cas7-11 (1,601 aa) enablespackaging along with a guide RNA cassette into single AAV viral vectorsfor delivery (FIG. 6E), with crRNA expression driven by either U6 ortRNA promoters. Testing Cas7-11S and ΔINS2 in HEK293FT cells via AAV8delivery (FIG. 6F), both variants were found to be able to effectivelyknock down Gluc mRNA, with higher knockdown efficiencies at highertiters and with tRNA-driven crRNA expression (FIGS. 6G and 20-23 ).

Example 16 Cas7-11 Function and Mechanism

The function and mechanism of Cas7-11 were assessed. The structure ofCas7-11 in complex with its mature crRNA and target RNA was observed(FIGS. 7A-7B). The structure observed at 2.5-Å reveals that the fourrepetitive Cas7 domains and Cas11 are interwoven via four interdomainlinkers (FIG. 7A). These linkers enable a multi-component type IIIsystem to emerge as a single-effector with all necessary domains encodedwithin the domains of a single protein and oriented correctly. The L4linker is unique as it weaves through the protein from Cas7.3 around theCTE domain and to Cas7.4 with twists that allow these domains to fittogether. The structure further reveals that Cas7.1 recognizes and bindsthe mature DR tag of the crRNA and is also containing the active sitefor pre-crRNA processing via an acid-base catalytic mechanism. Asprocessing in type III complexes is typically carried out by the enzymeCas6, this evolved function of type III-E effectors is unique and bearslittle resemblance to Cas6, allowing for Cas7-11 to function like otherClass 2 systems that have incorporated pre-crRNA processing like Cas12(e.g., (Swarts, van der Oost, and Jinek 2017), incorporated herewith inits entirety for reference) and Cas13 (e.g., East-Seletsky et al. 2016,incorporated herewith in its entirety for reference). Moreover, theprocessing domain of Cas7-11 (Cas7.1) has no resemblance to those ofCas12 (WED) and Cas13 (Helical-1), highlighting the mechanisticdiversity of pre-crRNA processing in CRISPR-Cas systems. Beyondprocessing, the structure also reveals how Cas7.2, Cas7.3, and Cas7.4bind the guide-target duplex and that through specific base flipping,Cas7.2 and Cas7.3 carry out precise cleavage 6-nt apart on the target,as in other type III systems like Csm3. By flipping out the bases of thetarget, Cas7-11 is able to cleave between the 3rd and 4th and 9th and10th nucleotides of the target by placing catalytic aspartate residuesin the vicinity of the scissile phosphodiester bonds. As other type IIIcomplexes typically have more Cas7 and Cas11 subunits than theequivalent domains of Cas7-11, they also have more cleavage sites withsome complexes cleaving up to 5 times (e.g., Mohanraju et al. 2016,incorporated herewith in its entirety for reference) (FIG. 7B).

One skilled in the art will appreciate further features and advantagesof the disclosure based on the above-described embodiments. Accordingly,the disclosure is not to be limited by what has been particularly shownand described, except as indicated by the appended claims.

Example 17 Single Mutant G-Luciferase Knockdown

Knockdown readout to measure cleavage activity on transcripts ofluminescent Guassia luciferase protein by wildtype and mutant DiCas7-11endonucleases was performed.

Sequences of mutant DiCas7-11 endonuclease coding sequences are shownbelow in Table 7 (short version of sequences showing in uppercase theresidues that are responsible to encode mutations) and Table 8 (completeversion of sequences).

TABLE 7 Residues responsible for mutation of DiCas7-11 SEQ IDResidues responsible for mutation of DiCas7-11 NO/Descriptionendonuclease coding sequences SEQ ID NO: 39gataataagaaaaccAAGtatacccgcctgcttg E279K SEQ ID NO: 40gaaaaccgaaAAGacccgcctgc Y280K SEQ ID NO: 41 tgccaaaggatAAGgacgggaaggaH305K SEQ ID NO: 42 gccaaaggatcatAAGgggaaggatgacc D306K SEQ ID NO: 43gacgggaaggatAAGcattacctctggg D310K SEQ ID NO: 44gaaggatgaccatAAGctctgggatatcgg Y312K SEQ ID NO: 45ggtgaagcgctgAAGctcaagagtaaa Y359K SEQ ID NO: 46 cggttgcgAAGggtgccctE512K SEQ ID NO: 47 tggcggatgAAGgctgatcacc D979K SEQ ID NO: 48cggatggatgctAAGcaccagaatgtg D981K SEQ ID NO: 49atgtgctgcaaAAGtttctcccaggt D988K SEQ ID NO: 50tagtgacaagaaagggAAGgtgatcaataactttca D1163K SEQ ID NO: 51tcagggtctataacAAGaatcctcaagcagt N1246K SEQ ID NO: 52tcattgttgAAGcggcctcgcc E1391K SEQ ID NO: 53 acaaagccctAAGaaccgcactgtN1440K SEQ ID NO: 54 gctgcgaaagaagAAGgatcctaatggga D1567K SEQ ID NO: 55acgaagaactcaaaAAGggggaattcaagaa D1580K SEQ ID NO: 56aataagaaaaccAGGtatacccgcctgc E279R SEQ ID NO: 57 aaaaccgaaAGGacccgcctgcY280R SEQ ID NO: 58 gccaaaggatAGGgacgggaagg H305R SEQ ID NO: 59ccaaaggatcatAGGgggaaggatgacc D306R SEQ ID NO: 60acgggaaggatAGGcattacctctgg D310R SEQ ID NO: 61aaggatgaccatAGGctctgggatatcg Y312R SEQ ID NO: 62gtgaagcgctgAGGctcaagagtaa Y359R SEQ ID NO: 63 ggttgcgAGGggtgccct E512RSEQ ID NO: 64 tggcggatgAGGgctgatcac D979R SEQ ID NO: 65ggatggatgctAGGcaccagaatgtg D981R SEQ ID NO: 66 tgtgctgcaaAGGtttctcccaggD988R SEQ ID NO: 67 agtgacaagaaagggAGGgtgatcaataacttt D1163RSEQ ID NO: 68 cagggtctataacAGGaatcctcaagcagt N1246R SEQ ID NO: 69cattgttgAGGcggcctcgc E1391R SEQ ID NO: 70 acaaagccctAGGaaccgcactg N1440RSEQ ID NO: 71 ctgcgaaagaagAGGgatcctaatggga D1567R SEQ ID NO: 72cgaagaactcaaaAGAggggaattcaaga D1580R

TABLE 8 SEQ ID NO/ DescriptionMutant DiCas7-11 endonuclease coding sequences SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA IDATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA NO: 119GCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACAT E279KAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCAAGTATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 120ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA Y280KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAAAAGACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 121ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA H305K GCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATAAGGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 122ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D306KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATAAGGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 123ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D310KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATAAGCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 124ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA Y312KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATAAGCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 125ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA Y359KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGAAGCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 126ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA E512KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGAAGGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 127ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D979KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGAAGGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 128ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D981KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTAAGCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 129ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D988KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAAAGTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 130ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D1163KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGAAGGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 131ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA N1246KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAAGAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 132ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA E1391KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTAT CTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGAAGCGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 133ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA N1440KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAAGAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 134ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D1567KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGAAGGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 135ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D1580KGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAAAGGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA NO: 136ATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA E279RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCAGGTATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 137ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA Y280RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAAAGGACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 138ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA H305RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATAGGGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 139ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D306RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATAGGGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA  ID NO: 140ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D310RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATAGGCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGA CTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 141ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA Y312RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATAGGCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 142ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA Y359RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGAGGCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 143ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA E512RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGAGGGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA ID NO: 144ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D979RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGAGGGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 145ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D981RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTAGGCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 146ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D988RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAAGGTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 147ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D1163RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGAGGGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 148ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA N1246RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAGGAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 149ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA E1391RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGAGGCGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 150ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA N1440RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAGGAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ ID ATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA NO: 151ATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAA D1567RGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGAGGGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAGACGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG SEQ IDACCGAATATACCCGCCTGCTTGCTGATGCTATTCGTTCACTCCGCCGCTCTAGTAA NO: 152ATGACGACTACTATGAAGATTTCAATTGAATTCCTCGAGCCGTTTCGGATGACCAA D1580RATGGCAGGAAAGCACTAGGCGAAACAAGAACAACAAGGAATTTGTTCGCGGCCAAGCTTTTGCAAGATGGCATAGAAATAAGAAGGATAACACAAAAGGGCGGCCTTACATAACTGGGACATTGCTGCGGTCCGCAGTAATTAGAAGCGCGGAAAACCTGCTGACATTGAGTGATGGGAAAATTAGCGAAAAGACATGTTGTCCTGGGAAATTTGATACGGAAGATAAAGACAGGTTGCTGCAATTGAGGCAGCGGTCCACCCTTCGTTGGACCGATAAGAACCCCTGTCCAGATAACGCTGAAACTTACTGCCCGTTTTGCGAACTTCTCGGGAGATCCGGTAACGATGGAAAGAAGGCTGAGAAGAAGGATTGGCGCTTTCGTATTCATTTTGGCAATCTTTCCCTTCCAGGGAAGCCCGATTTCGACGGGCCAAAAGCTATAGGCAGCCAACGGGTACTTAACAGGGTCGATTTCAAATCCGGTAAGGCCCATGACTTCTTTAAGGCTTACGAAGTCGACCATACTAGGTTTCCCCGCTTCGAAGGGGAGATTACCATAGATAATAAAGTCAGCGCTGAAGCCAGGAAACTGCTCTGTGATTCTCTCAAGTTTACGGATCGGCTGTGTGGAGCTCTGTGCGTAATCAGGTTCGATGAATATACACCAGCAGCCGATAGTGGGAAACAAACCGAAAATGTCCAAGCAGAACCGAATGCTAATCTCGCTGAAAAGACCGCAGAGCAAATTATTAGCATTCTGGACGATAATAAGAAAACTTGTCGCTGGCTTGCCAAAGGATCATGACGGGAAGGATGACCATTACCTCTGGGATATCGGCAAGAAGAAGAAAGACGAAAACTCTGTCACTATTAGGCAAATCCTTACGACCTCAGCAGACACCAAGGAACTCAAGAATGCCGGAAAATGGAGAGAATTCTGCGAGAAGCTGGGTGAAGCGCTGTACCTCAAGAGTAAAGACATGAGTGGCGGCCTGAAAATTACTCGGAGAATACTGGGCGATGCTGAATTCCATGGAAAGCCCGATCGGCTTGAAAAGAGCCGGTCTGTGTCAATCGGATCTGTGTTGAAAGAAACTGTGGTATGCGGCGAACTGGTCGCTAAAACGCCCTTCTTCTTCGGAGCGATAGACGAAGATGCAAAACAAACCgacCTGCAAGTACTCCTCACTCCCGATAACAAGTATAGACTGCCAAGAAGCGCCGTGCGAGGTATACTCCGTCGAGATCTTCAAACCTATTTTGATAGCCCATGCAATGCTGAGTTGGGTGGACGGCCATGCATGTGTAAGACGTGTAGGATTATGAGAGGGATCACGGTGATGGATGCGCGCAGTGAGTACAATGCCCCGCCAGAAATAAGGCATCGTACCCGCATTAATCCCTTCACAGGCACGGTTGCGGAAGGTGCCCTGTTTAATATGGAAGTAGCCCCCGAGGGGATTGTCTTTCCATTTCAACTCCGGTACCGGGGCTCTGAAGATGGGCTGCCCGATGCACTGAAAACGGTGTTGAAATGGTGGGCTGAGGGGCAGGCATTCATGAGTGGCGCTGCCTCAACCGGGAAGGGCCGATTCCGGATGGAAAATGCTAAATATGAAACGCTGGATCTGAGCGACGAGAATCAAAGGAATGACTATCTTAAGAATTGGGGATGGCGTGACGAGAAGGGGCTCGAGGAACTGAAGAAACGACTGAACTCAGGTCTGCCAGAGCCCGGTAATTATAGGGATCCAAAATGGCACGAGATTAACGTTTCCATTGAGATGGCAAGCCCTTTTATTAATGGCGACCCAATCCGCGCAGCCGTGGACAAACGTGGTACAgatGTGGTTACCTTCGTTAAGTATAAAGCTGAAGGGGAAGAGGCGAAACCCGTATGTGCATACAAGGCCGAATCTTTTAGAGGGGTGATCAGAAGTGCCGTGGCACGCATTCATATGGAAGATGGCGTCCCTTTGACTGAGTTGACTCACAGTGACTGTGAATGTCTCCTGTGCCAAATCTTTGGAAGTGAGTATGAAGCCGGCAAAATAAGGTTTGAAGATCTCGTATTCGAAAGTGACCCGGAACCTGTGACCTTCGATCATGTGGCCATCGATAGATTCACTGGCGGTGCAGCTGATAAGAAGAAATTCGATGATTCCCCTCTGCCCGGTAGCCCTGCAAGACCGCTCATGTTGAAAGGCTCCTTCTGGATCCGCAGGGACGTTCTCGAGGACGAAGAGTACTGTAAGGCACTCGGTAAGGCTCTTGCAGATGTGAATAATGGCCTTTATCCCCTCGGTGGAAAGAGCGCCATCGGCTACGGACAGGTCAAGAGTCTGGGTATAAAGGGAGATGATAAGAGGATTTCTCGCCTCATGAATCCTGCCTTTGATGAGACAGATGTAGCCGTTCCAGAAAAGCCCAAAACTGATGCCGAGGTTCGCATCGAGGCAGAGAAAGTATATTACCCACACTATTTCGTCGAACCCCATAAGAAGGTGGAACGCGAGGAGAAACCCTGTGGTCATCAAAAGTTCCACGAGGGGCGACTGACAGGTAAAATTCGGTGTAAGCTCATTACCAAGACACCCCTCATCGTCCCAGATACTAGTAATGACGATTTCTTCAGACCTGCGGATAAAGAAGCTCGGAAGGAAAAGGACGAATATCATAAATCATATGCTTTCTTCAGACTTCATAAACAAATCATGATTCCCGGGAGCGAATTGAGAGGAATGGTGAGTAGTGTCTACGAAACTGTGACAAATTCTTGCTTCAGGATATTTGATGAGACTAAACGGTTGTCATGGCGGATGGATGCTGATCACCAGAATGTGCTGCAAGACTTTCTCCCAGGTCGAGTGACCGCCGATGGGAAACATATACAAAAGTTTTCCGAAACTGCAAGGGTGCCTTTCTATGACAAAACGCAGAAACATTTTGACATTCTTGATGAGCAAGAAATTGCTGGTGAGAAACCTGTTCGGATGTGGGTCAAGCGTTTTATTAAACGACTGAGCCTCGTTGATCCTGCTAAACACCCCCAGAAGAAACAAGACAATAAATGGAAAAGACGCAAAGAAGGCATCGCCACATTTATAGAGCAAAAGAATGGCTCTTATTATTTTAACGTGGTCACCAATAATGGCTGCACTTCTTTCCACTTGTGGCATAAACCTGACAATTTTGACCAGGAGAAACTCGAAGGCATACAGAATGGTGAGAAACTGGATTGCTGGGTAAGAGATAGTAGATACCAAAAGGCCTTTCAAGAGATACCCGAGAACGACCCAGACGGATGGGAGTGTAAAGAGGGCTACCTTCATGTCGTCGGCCCCAGCAAAGTAGAGTTTAGTGACAAGAAAGGGGATGTGATCAATAACTTTCAAGGAACACTCCCATCAGTTCCCAACGACTGGAAAACAATTAGGACGAACGATTTTAAGAATAGGAAAAGGAAGAATGAACCTGTGTTTTGTTGTGAGGATGATAAGGGCAATTACTATACCATGGCTAAATATTGCGAAACCTTCTTCTTCGATCTGAAGGAGAACGAGGAATACGAGATCCCCGAGAAAGCAAGAATCAAATACAAAGAACTGCTCAGGGTCTATAACAACAATCCTCAAGCAGTGCCGGAGAGCGTATTTCAGTCTAGAGTTGCCCGGGAAAACGTGGAAAAGCTGAAGTCCGGAGATCTTGTGTATTTCAAACATAATGAAAAGTACGTAGAGGACATCGTCCCAGTGCGGATTTCCCGAACTGTAGACGATAGGATGATCGGCAAACGTATGAGCGCCGATCTGCGGCCGTGCCATGGAGATTGGGTGGAAGATGGTGATCTCAGTGCCTTGAATGCATATCCCGAGAAAAGACTCCTCTTGCGCCACCCCAAAGGACTCTGCCCTGCTTGCCGGCTCTTTGGAACCGGATCTTACAAGGGCAGAGTCAGGTTTGGATTCGCGTCACTCGAAAACGATCCGGAGTGGCTGATCCCAGGCAAGAATCCCGGCGATCCGTTTCACGGCGGGCCGGTGATGCTCTCATTGTTGGAACGGCCTCGCCCGACTTGGAGTATACCGGGATCCGACAATAAGTTTAAAGTGCCTGGCAGAAAGTTTTACGTCCACCACCACGCCTGGAAAACCATTAAGGACGGGAACCATCCCACAACAGGCAAAGCTATTGAACAAAGCCCTAATAACCGCACTGTAGAAGCTCTCGCCGGCGGGAATTCCTTTAGCTTCGAAATTGCCTTTGAGAACCTGAAAGAATGGGAGCTGGGTTTGCTCATCCACAGCCTGCAACTCGAAAAGGGTCTGGCGCATAAACTTGGAATGGCAAAGTCTATGGGATTTGGTTCAGTTGAAATTGACGTCGAATCAGTGCGCCTGAGAAAAGATTGGAAGCAATGGCGGAATGGCAATTCCGAAATTCCCAACTGGTTGGGAAAAGGATTTGCTAAACTGAAGGAATGGTTCCGGGACGAGCTCGATTTTATAGAAAATCTTAAGAAACTTCTTTGGTTTCCTGAGGGCGACCAAGCACCCCGGGTTTGCTACCCCATGCTGCGAAAGAAGGACGATCCTAATGGGAATAGCGGTTACGAAGAACTCAAAAGGGGGGAATTCAAGAAAGAAGATCGGCAGAAGAAGCTGACCACGCCGTGGACACCGTGGGCATAG

Mutants were nominated from the solved structure of Desulfonemaishimotonii and generated by site directed mutagenesis with primers,then assembled by Gibson assembly. Cloned variants were transformed intoE. coli, grown overnight, then picked into TB media for outgrowth.Following an outgrowth period, a Qiagen 96well-miniprep protocol wasused to purify plasmid and correct cloning was confirmed through Tn5fragmentation and sequencing on an Illumina MiSeq. All constructs weretransfected at a 96-well scale on HEK293FT cells in DMEM 10% FBS, alongwith a guide targeting the g-luciferase transcript (SEQ ID NO:73,sequence 5′-TGCAGCCAGCTITCCGGGCATTGGCTTCCAT-3′) and a reporter plasmidexpressing Gaussia-luciferase and Cypridina luciferase. For allknockdown experiments, 10 ng of guide, 10 ng of target plasmid, and 40ng of DiCas7-11 plasmids were co-transfected using Lipofectamine 3000.

G-luciferase levels were read using a 96-well plate reader 48 hpost-transfection and normalized against C-luciferase levels. Results ofthe readout are shown in FIG. 25 .

Endonuclease constructs are shown across the x-axis, the y-axis displaysthe relative G-luciferase to C-luciferase level for each construct,normalized to a non-targeting guide (SEQ ID NO:74, sequence5′-GGTAATGCCTGGCTTGTCGACGCATAGTCTG-3′).

Example 18 Single Mutant Endogenous MALAT1 Knockdown

Knockdown readout to measure cleavage activity on endogenous MALAT1transcripts by wildtype and mutant DiCas7-11 endonucleases wasperformed.

Mutants were nominated from the solved structure of Desulfonemaishimotonii and generated by site directed mutagenesis with primers,then assembled by Gibson assembly. Cloned variants were transformed intoE. coli, grown overnight, then picked into TB media for outgrowth.Following an outgrowth period, a Qiagen 96well-miniprep protocol wasused to purify plasmid and correct cloning was confirmed through Tn5fragmentation and sequencing on an Illumina MiSeq. All constructs weretransfected at a 96-well scale on HEK293FT cells in DMEM 10% FBS, alongwith a guide targeting MALAT1 (SEQ ID NO:75, sequence5′-GGTTATAGCTGACAAGCAATTAACTTAAA-3′). Knockdown transfections combine 10ng of guide and 40 ng of DiCas7-11 plasmids and are performed usingLipofectamine 3000. RNA was harvested 3 days post-transfection andreverse transcribed using the Thermo RevertAid cDNA prep kid with randomhexamer and poly-dT primers. qPCR was performed on all samples usingassays targeting endogenous MALAT and GAPDH as a control.

Results of the readout are shown in FIG. 26 . Endonuclease constructsare shown across the x-axis, the y-axis displays the relative MALATexpression with each mutant normalized to a non-targeting guide (SEQ IDNO:74, sequence 5′-GGTAATGCCTGGCTTGTCG ACGCATAGTCTG-3′).

Example 19 Single Mutant Endogenous PPIB Knockdown

Knockdown readout to measure cleavage activity on transcripts ofluminescent Guassia luciferase protein by wildtype and mutant DiCas7-11endonucleases was performed.

Mutants were nominated from the solved structure of Desulfonemaishimotonii and generated by site directed mutagenesis with primers,then assembled by Gibson assembly. Cloned variants were transformed intoE. coli, grown overnight, then picked into TB media for outgrowth.Following an outgrowth period, a Qiagen 96well-miniprep protocol wasused to purify plasmid and correct cloning was confirmed through Tn5fragmentation and sequencing on an Illumina MiSeq. All constructs weretransfected at a 96-well scale on HEK293FT cells in DMEM 10% FBS, alongwith a guide targeting PPIB (SEQ ID NO:76, sequence5′-cagtgttggtaggagtttgttacaaaagtga-3′). Knockdown transfections combine10 ng of guide and 40 ng of DiCas7-11 plasmids and are performed usingLipofectamine 3000. RNA was harvested 3 days post-transfection andreverse transcribed using the Thermo RevertAid cDNA prep kid with randomhexamer and poly-dT primers. qPCR was performed on all samples usingassays targeting endogenous PPIB and GAPDH as a control.

Results of the readout are shown in FIG. 27 . Endonuclease constructsare shown across the x-axis, the y-axis displays the relative PPIBexpression with each mutant normalized to a non-targeting guide (SEQ IDNO:74, sequence 5′-GGTAATGCCTGGCTTGTCG ACGCATAGTCTG-3′).

Example 20 Single and Double Mutant G-Luciferase Knockdown

Knockdown readout to measure cleavage activity on transcripts ofluminescent Guassia luciferase protein by wildtype and mutant DiCas7-11endonucleases was performed.

Mutants were nominated from the solved structure of Desulfonemaishimotonii and generated by site directed mutagenesis with primers,then assembled by Gibson assembly. Cloned variants were transformed intoE. coli, grown overnight, then picked into TB media for outgrowth.Following an outgrowth period, a Qiagen 96well-miniprep protocol wasused to purify plasmid and correct cloning was confirmed through Tn5fragmentation and sequencing on an Illumina MiSeq. Double mutants shownhere were created by cloning combinations of working mutations using thesame method employed for the 1st round hits. All constructs weretransfected at a 96-well scale on HEK293FT cells in DMEM 10% FBS, alongwith a guide targeting the g-luciferase transcript (SEQ ID NO:73,sequence 5′-TGCAGCCAGCTITCCGGGCATTGGCTTCCAT-3′) and a reporter plasmidexpressing Gaussia-luciferase and Cypridina luciferase. For allknockdown experiments, 10 ng of guide, 10 ng of target plasmid, and 40ng of DiCas7-11 plasmids were co-transfected using Lipofectamine 3000.G-luciferase levels were read using a 96-well plate reader 48 hpost-transfection and normalized against C-luciferase levels.

Results of the readout are shown in FIG. 28 . Endonuclease constructsare shown across the x-axis, the y-axis displays the relativeG-luciferase to C-luciferase level for each construct, normalized to anon-targeting guide (SEQ ID NO:74, sequence 5′-GGTAATGCCTGGCTTGTCGACGCATAGTCTG-3′).

Example 21 Saturation Mutagenesis for D1580 Residue

Knockdown readout to measure cleavage activity on transcripts ofluminescent Guassia luciferase protein by wildtype and mutant DiCas7-11endonucleases was performed.

Mutants generated by site directed mutagenesis for the amino acidresidue D1580, which was substituted by each other residue. Thesmall-discas7-11 constructs also shown here are based on the 2021structure paper from Kato et al. (Kato K, Zhou W, Okazaki S, Isayama Y,Nishizawa T, Gootenberg J S, Abudayyeh O O, Nishimasu H. Structure andengineering of the type III-E CRISPR-Cas7-11 effector complex. Cell.2022 Jun. 23; 185(13):2324-2337.e16. doi: 10.1016/j.cell.2022.05.003.Epub 2022 May 27. PMID: 35643083) which replaced the Cas7-11 INS domainwith a short GS linker sequence. Mutations were also generated on top ofthe small-cas scaffold. Cloned variants were transformed into E. coli,grown overnight, then picked into TB media for outgrowth. Following anoutgrowth period, a Qiagen 96well-miniprep protocol was used to purifyplasmid and correct cloning was confirmed through Tn5 fragmentation andsequencing on an Illumina MiSeq. All constructs were transfected at a96-well scale on HEK293FT cells in DMEM 10% FBS, along with a guidetargeting the g-luciferase transcript (SEQ ID NO:73, sequence5′-TGCAGCCAGCTITCCGGGCATTGGCTTCCAT-3′) and a reporter plasmid expressingGaussia-luciferase and Cypridina luciferase. For all knockdownexperiments, 10 ng of guide 10 ng of target plasmid and 40 ng ofDiCas7-11 plasmids were co-transfected using Lipofectamine 3000.G-luciferase levels were read using a 96-well plate reader 48 hpost-transfection and normalized against C-luciferase levels.

Results of the readout are shown in FIG. 29 . Endonuclease constructsare shown across the x-axis, the y-axis displays the relativeG-luciferase to C-luciferase level for each construct, normalized to anon-targeting guide (SEQ ID NO:74, sequence 5′-GGTAATGCCTGGCTTGTCGACGCATAGTCTG-3′).

Example 22 Single, Double, Triple, and Quadrupole Mutants G-LuciferaseKnockdown

Knockdown readout to measure cleavage activity on transcripts ofluminescent Guassia luciferase protein by wildtype and mutant DiCas7-11endonucleases was performed.

Mutants generated by site directed mutagenesis for the amino acidresidue D1580, which was substituted by each other residue. Thesmall-discas7-11 constructs also shown here are based on the 2021structure paper from Kato et al. (Kato K, Zhou W, Okazaki S, Isayama Y,Nishizawa T, Gootenberg J S, Abudayyeh O O, Nishimasu H. Structure andengineering of the type III-E CRISPR-Cas7-11 effector complex. Cell.2022 Jun. 23; 185(13):2324-2337.e16. doi: 10.1016/j.cell.2022.05.003.Epub 2022 May 27. PMID: 35643083) which replaced the DiCas7-11 INSdomain with a short GS linker sequence. Mutations were also generated ontop of the small-cas scaffold. Cloned variants were transformed into E.coli, grown overnight, then picked into TB media for outgrowth.Following an outgrowth period, a Qiagen 96well-miniprep protocol wasused to purify plasmid and correct cloning was confirmed through Tn5fragmentation and sequencing on an Illumina MiSeq. Triple and quadruplemutants (residues as listed) were generated using the same PCRmutagenesis strategy employed for the single and double mutants. Allconstructs were transfected at a 96-well scale on HEK293FT cells in DMEM10% FBS, along with a guide targeting the g-luciferase transcript (SEQID NO:73, sequence 5′-TGCAGCCAGCTITCCGGGCATTGGCTTCCAT-3′) and a reporterplasmid expressing Gaussia-luciferase and Cypridina luciferase. For allknockdown experiments, 10 ng of guide, 10 ng of target plasmid, and 40ng of DiCas7-11 plasmids were co-transfected using Lipofectamine 3000.G-luciferase levels were read using a 96-well plate reader 48 hpost-transfection and normalized against C-luciferase levels.

Results of the readout are shown in FIG. 30 . Endonuclease constructsare shown across the x-axis, the y-axis displays the relativeG-luciferase to C-luciferase level for each construct, normalized to anon-targeting guide (SEQ ID NO:74, sequence 5′-GGTAATGCCTGGCTTGTCGACGCATAGTCTG-3′).

All references (e.g., publications or patents or patent applications)cited herein are incorporated herein by reference in their entiretiesand for all purposes to the same extent as if each individual reference(e.g., publication or patent or patent application) was specifically andindividually indicated to be incorporated by reference in its entiretyfor all purposes.

LIST OF REFERENCES

All publications and references cited herein are expressly incorporatedherein by reference in their entirety.

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Kato K, Zhou W, Okazaki S, Isayama Y, Nishizawa T, Gootenberg J S,Abudayyeh O O, Nishimasu H. Structure and engineering of the type III-ECRISPR-Cas7-11 effector complex. Cell. 2022 Jun. 23;185(13):2324-2337.e16. doi: 10.1016/j.cell.2022.05.003. Epub 2022 May27. PMID: 35643083

1. A genome editing system capable of cleaving an RNA target, the systemcomprising: a polypeptide comprising an amino acid sequence at least 85%identical to the amino acid sequence of any one of SEQ ID NOs: 1-4wherein the amino acid sequence of the polypeptide comprises at leastone amino acid modification or mutation relative to the amino acidsequence of SEQ ID NO: 1-4, a deaminase, and a guide RNA that hybridizesto the RNA target and the polypeptide. 2-6. (canceled)
 7. The genomeediting system of claim 1, wherein the at least one amino acidmodification or mutation comprises: removing an amino acid; adding anamino acid; replacing an amino acid with no charge with an amino acidwith a positive charge; or replacing an amino acid with a negativecharge with an amino acid with a positive charge.
 8. The genome editingsystem of claim 7, wherein: the amino acid without charge is selectedfrom the group consisting of serine, threonine, asparagine, glutamine,cysteine, glycine, proline, alanine, valine, isoleucine, leucine,methionine, phenylalanine, tyrosine, and tryptophan; the amino acid witha negative charge is selected from the group consisting of aspartic acidand glutamic acid: or the amino acid with a positive charge is selectedfrom the group consisting of arginine, histidine, and lysine. 9-10.(canceled)
 11. The genome editing system of claim 1, wherein the aminoacid sequence of the polypeptide comprises 1, 2, 3, or 4 amino acidmodifications or mutations.
 12. The genome editing system of claim 1,wherein the amino acid sequence of the polypeptide comprises an alanineat a position corresponding to position 43 of SEQ ID NO: 1; an alanineat a position corresponding to position 55 of SEQ ID NO: 55; and/or analanine at a position corresponding to position 152 of SEQ ID NO:
 1. 13.The genome editing system of claim 1, wherein the polypeptide comprisesa deletion of one or more amino acid residues at positions 979 through1293 of SEQ ID NO: 1; at positions 1007 through 1220 of SEQ ID NO: 1;and/or at positions 1146 through 1211 of SEQ ID NO:
 1. 14. A compositionthat cleaves an RNA target comprising: a polypeptide comprising an aminoacid sequence at least 85% identical to the amino acid sequence of anyone of SEO ID NOs: 1-4 wherein the amino acid sequence of thepolypeptide comprises at least one amino acid modification or mutationrelative to the amino acid sequence of SEO ID NO: 1-4, a deaminase, anda guide RNA that hybridizes to the RNA target and the polypeptide. 15.The composition of claim 14, wherein: the guide RNA comprises a mismatchdistance that is about 20-65% of the length of the guide; the guide RNAhas a sequence with a length of about 20 to about 53 nucleotides (nt),optionally about 25 to about 53 nt, optionally about 29 to about 53 nt,or optionally about 40 to about 50 nt; the guide RNA is a pre-crRNA; theguide RNA is a mature crRNA; the RNA target is a single-strand RNA(ssRNA); the RNA target is in a cell, optionally wherein the cell is aprokaryotic cell or eukaryotic cell, optionally wherein the eukaryoticcell is a mammalian cell, optionally wherein the mammalian cell is ahuman cell; or the guide RNA comprises a mismatch that is about 20 toabout 30 nucleotides from a non-pairing C of the guide RNA. 16-25.(canceled)
 26. One or more nucleic acid molecules encoding thepolypeptide, the deaminase, and the guide RNA of claim
 14. 27-28.(canceled)
 29. A vector comprising the one of more nucleic acidmolecules of claim 26, wherein the vector is a viral vector, optionallywherein the viral vector is a lenti-associated viral vector,baculo-associated viral vector, or adeno-associated viral vector, andoptionally wherein the viral vector is derived from a virus selectedfrom the group consisting of Myoviridae, Siphoviridae, Podoviridae,Corticoviridae, Lipothrixviridae, Poxviridae, Iridoviridae,Adenoviridae, Polyomaviridae, Papillomaviridae, Mimiviridae,Pandoravirusa, Salterprovirusa, Inoviridae, Microviridae, Parvoviridae,Circoviridae, Hepadnaviridae, Caulimoviridae, Retroviridae,Cystoviridae, Reoviridae, Birnaviridae, Totiviridae, Partitiviridae,Filoviridae, Orthomyxoviridae, Deltavirusa, Leviviridae, Picornaviridae,Marnaviridae, Secoviridae, Potyviridae, Caliciviridae, Hepeviridae,Astroviridae, Nodaviridae, Tetraviridae, Luteoviridae, Tombusviridae,Coronaviridae, Arteriviridae, Flaviviridae, Togaviridae, Virgaviridae,Bromoviridae, Tymoviridae, Alphaflexiviridae, Sobemovirusa, Idaeovirusa,and Herpesviridae.
 30. A cell comprising the composition of claim 14,optionally wherein: the cell is a prokaryotic cell, or the cell is aeukaryotic cell, optionally wherein the eukaryotic cell is a mammaliancell, optionally wherein the mammalian cell is a human cell. 31-34.(canceled)
 35. A method of cleaving an RNA target in a cell, stabilizingan RNA target in a cell, or affecting translation of an RNA target in acell, the method comprising providing to the cell the composition ofclaim 14, optionally wherein the RNA target is an ssRNA. 36-38.(canceled)
 39. A method of treating a genetically inherited disease in asubject in need thereof comprising administering to the subject aneffective amount of the composition of claim 14, wherein the geneticallyinherited disease involves a guanosine to adenosine change in a genomeof the subject or is a pre-termination disease.
 40. The method of claim39, wherein the genetically inherited disease is selected from the groupconsisting of Meier-Gorlin syndrome; Seckel syndrome 4; Joubert syndrome5; Leber congenital amaurosis 10; Charcot-Marie-Tooth disease, type 2;leukoencephalopathy; Usher syndrome, type 2C; spinocerebellar ataxia 28;glycogen storage disease type III; primary hyperoxaluria, type I; longQT syndrome 2; Sjögren-Larsson syndrome; hereditary fructosuria;neuroblastoma; amyotrophic lateral sclerosis type 9; Kallmann syndrome1; limb-girdle muscular dystrophy, type 2L; familial adenomatouspolyposis 1; familial type 3 hyperlipoproteinemia; Alzheimer's disease,type 1; metachromatic leukodystrophy; cancer; Uveitis; SCA1; SCA2;FUS-Amyotrophic Lateral Sclerosis (ALS); MAPT-Frontotemporal Dementia(FTD); Myotonic Dystrophy Type 1 (DM1); Diabetic Retinopathy (DR/DME);Oculopharyngeal Muscular Dystrophy (OPMD); SCA8; C90RF72-AmyotrophicLateral Sclerosis (ALS); SOD1-Amyotrophic Lateral Sclerosis (ALS);Spinal Cord Injury (targets: mTOR, PTEN, KLF6/7, SOX11, KCC2, and growthfactors); SCA6; SCA3 (Machado-Joseph Disease); Multiple system Atrophy(MSA); Treatment-resistant Hypertension; Myotonic Dystrophy Type 2(DM2); Fragile X-associated Tremor Ataxia Syndrome (FXTAS); WestSyndrome with ARX Mutation; Age-related Macular Degeneration(AMD)/Geographic Atrophy (GA); C90RF72-Frontotemporal Dementia (FTD);Facioscapulohumeral Muscular Dystrophy (FSHD); Fragile X Syndrome (FXS);Huntington's Disease; Glaucoma; Acromegaly; Achromatopsia (total colorblindness); Ullrich congenital muscular dystrophy; Hereditary myopathywith lactic acidosis; X-linked spondyloepiphyseal dysplasia tarda;Neuropathic pain (Target: CPEB); Persistent Inflammation and injury pain(Target: PABP); Neuropathic pain (Target: miR-30c-5p); Neuropathic pain(Target: miR-195); Friedreich's Ataxia; Uncontrolled gout; Inflammatorypain (Target: Nav1.7 and Nav1.8); Choroideremia; Focal epilepsy; Alpha-1Antitrypsin deficiency (AATD); Androgen Insensitivity Syndrome;Opioid-induced hyperalgesia (Target: Raf-1); Neurofibromatosis type 1;Stargardt's Disease; Dravet Syndrome; Retinitis Pigmentosa; andParkinson's Disease.
 41. (canceled)
 42. A method of altering splicing ofa pre-mRNA in a cell, increasing RNA stability in a cell, or modulatingtranslation in a cell, the method comprising administering to the cellan effective amount of the composition of claim
 14. 43. A method ofchanging microRNA targets in a subject in need thereof comprisingadministering to the subject an effective amount of the composition ofclaim
 14. 44-45. (canceled)
 46. A method of detecting a bacterium orderivative thereof in a sample, the method comprising: adding to thesample an effective amount of the composition of claim 14; and detectinga reporter specific to the bacterium or derivative thereof.
 47. A methodof detecting a virus or derivative thereof in a sample, the methodcomprising: adding to the sample an effective amount of the compositionsof claim 14; and detecting a reporter specific to the virus orderivative thereof.
 48. The genome editing system of claim 1, whereinthe deaminase is selected from the group consisting of an adenosinedeaminase or a catalytic domain thereof, and a cytidine deaminase or acatalytic domain thereof.
 49. A genome editing system capable ofcleaving an RNA target, the system comprising: a polypeptide comprisingan amino acid sequence at least 85% identical to the amino acid sequenceof any one of SEQ ID NOs: 1-4, wherein the polypeptide comprises adeaminase that is fused or linked to the polypeptide, and wherein theamino acid sequence of the polypeptide comprises at least one amino acidmodification or mutation relative to the amino acid sequence of SEQ IDNO: 1-4, and a guide RNA that hybridizes to the RNA target and thepolypeptide.